LOCUS       sequence-134594-        5532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Floxed Kanamycin (G418) selection maker; 3' tagging vector used to 
            create C-terminal fusion proteins with 1x-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5532)
  AUTHORS   Saroufim MA, Bensidoun P, Raymond P, Rahman S, Krause MR, Oeffinger 
            M, Zenklusen D
  TITLE     The nuclear basket mediates perinuclear mRNA scanning in budding 
            yeast.
  JOURNAL   J Cell Biol. 2015 Dec 21;211(6):1131-40. doi: 10.1083/jcb.201503070.
  PUBMED    26694838
REFERENCE   2  (bases 1 to 5532)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2015-12-21- 21"; volume: "211"; 
            issue: "6"; pages: "1131-40"
COMMENT     Sequence Label: pDZ584 pKAN 1x-mCherry
FEATURES             Location/Qualifiers
     source          1..5532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      11..20
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             17..727
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(162..180)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     430..449
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(748..764)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(749..765)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      767..1014
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(783..801)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    1065..1098
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            1153..2509
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1153..1496
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1497..2306
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1564..1583)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2312..2509
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2572..2605
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2656..2676)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2656..2674)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2695..2711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2695..2711)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2708..2730)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     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)."
     promoter        complement(join(2743..2749,2750..2767,2768..2773))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2788..2809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2926..2943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3097..3685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3177..3196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3856..4647,4648..4716))
                     /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     4479..4498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4717..4821)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4848..5303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4935..4954)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5145..5166
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5429..5451
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5443..5460
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5444..5460
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5470..5489
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5470..5488
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(5521..5532,1..5)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5521..5532,1..5)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ggatccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
       61 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      121 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      181 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
      241 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
      301 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
      361 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
      421 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
      481 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
      541 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
      601 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
      661 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
      721 caagtaagaa ttcgatatca agcttatcga taccgtcgac ctcgagtcat gtaattagtt
      781 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
      841 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
      901 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
      961 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg
     1021 tacccagctg aagcttcgta cgctgcaggt cgacaaccct taatataact tcgtataatg
     1081 tatgctatac gaagttatta ggtctagaga tctgtttagc ttgcctcgtc cccgccgggt
     1141 cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc
     1201 tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca
     1261 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
     1321 cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
     1381 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
     1441 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg
     1501 gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc caacatggat gctgatttat
     1561 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     1621 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1681 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1741 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggca
     1801 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1861 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1921 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1981 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     2041 agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2101 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     2161 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2221 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2281 atttgatgct cgatgagttt ttctaatcag tactgacaat aaaaagattc ttgttttcaa
     2341 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
     2401 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     2461 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg
     2521 atactaacgc cgccatccag tgtcgaaaac gagctctcga gaacccttaa tataacttcg
     2581 tataatgtat gctatacgaa gttattaggt gatatcagat ccactagtgg cctatgcggt
     2641 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2701 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     4861 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4921 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4981 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5041 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5101 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5161 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5221 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5281 caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5341 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5401 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     5461 gagcgcgcgt aatacgactc actatagggc gaattggagc tccaccgcgg tggcggccgc
     5521 tctagaacta gt
//
