LOCUS       sequence-243552-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for tagging proteins in Cryptococcus neoformans; C-terminal 
            fragment of Venus protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5318)
  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 5318)
  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: YSBE415: pTOP_ACA1_Vc_HOG1 terminator
FEATURES             Location/Qualifiers
     source          1..5318
                     /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(614..865)
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(656..677)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1901..1918)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1901..1917)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1910..1932)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2055..2357
                     /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     2236..2255
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2706..3500
                     /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             2706..3500
                     /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(2760..2779)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3370..3389
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3518..3586,3587..4378)
                     /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(3736..3755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4549..5137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5038..5057
                     /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 ctacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
      661 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggt
      721 agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
      781 ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg
      841 ccttgatgcc gttcttctgc ttgtcagtac caccagaacc tcctgagtgc tctacaatct
      901 cggtaaccag ttttccgtcg attatccttg acttcatctg ctcaggcaca ggcctttcga
      961 cgaaatcacc atcttcgccg gtgggcacag aaatgttgat gctactggac ttgctggtaa
     1021 cgatttcaac cacttccata caatcctttg agaggtatac ctggccagag tcagtggtgt
     1081 cgatttggat ggtgggaatg gagccgatga tctggacttc aaaggagggg gacgaagtga
     1141 tagagaaaga ggatacggcg gtgtcgagga cgacagacgt cttcttacat ccagctgcga
     1201 gttatcagct atatgcatat gtgctaagag cattgactat acttacccat ggttacagcg
     1261 ttgattttac cggagatctg aaccacagag ttgacgcaac caaagatgtg aacagtgtgg
     1321 tgaagttcag tgttgtcgat tttaatagat ttgttgtcct cttggttttc ctgttataag
     1381 tcagtcacaa aacgagacca tgaaaggaca aagaaaagag acgaacaatg atccatttgt
     1441 tcccatcttc aagctcgatc ttcgcaggct tctttgcagg ctttgcccca ggcttgggtt
     1501 tgagaggcgg gcccttcttt gcattgtcag agacagtgcc ggcagaacga agtgaggggt
     1561 tcttgtgggt catctgggaa ctatcgacct ttcgcaatcc agaagtgaca gcgccaccac
     1621 gattcaaatc agccaaaagg gcagcagctc cagaaccaga tgtaggagca gccggtgctg
     1681 agccagaagg agggggagga ggtggagaag gagcagatgc tgatgccttg ggcatagaag
     1741 ggggggccgg ggaaccctgt gagagttagc agagttagcg acgctcggaa taagcagatg
     1801 gatccaaggg cgaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     1861 gagggcccaa ttcgccctat agtgaatcgt attacaattc actggccgtc gttttacaac
     1921 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1981 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2041 gcctatacgt acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     2101 ttgtggatgt acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg
     2161 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     2221 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
     2281 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     2341 tgttctgggg aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc
     2401 ttttcacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     2461 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     2521 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     2581 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     2641 cgccaaggat ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt
     2701 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2761 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2821 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2881 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     2941 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3001 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3061 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3121 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3181 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3241 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3301 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3361 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3421 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3481 ttcttgacga gttcttctga attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3721 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4201 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagcc aggcaactat
     4321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4801 taagtcgtgt cttaccgggt tggattcaag acgatagtta ccggataagg cgcagcggtc
     4861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5161 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5221 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5281 gaccgagcgc agcgaatcag tgagcgagga agcggaag
//
