LOCUS       sequence-131685-        5192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Floxed Kanamycin (G418) selection maker; 3' tagging vector used to 
            add 12 repetitive PP7 version 4 RNA stem-loops to gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  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 5192)
  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: pDZ617 pKAN 12xPP7 V4
FEATURES             Location/Qualifiers
     source          1..5192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..48
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     32..48
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    540..573
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      599..786
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(654..678)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            856..2212
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        856..1199
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1200..2009
                     /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(1267..1286)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2015..2212
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2275..2308
                     /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(2359..2379)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2359..2377)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2398..2414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2398..2414)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2411..2433)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2422..2438
                     /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(2446..2452,2453..2470,2471..2476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2491..2512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2629..2646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2800..3388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2880..2899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3559..4350,4351..4419))
                     /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     4182..4201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4420..4524)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4551..5006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4638..4657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4848..4869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5132..5154
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5146..5163
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5147..5163
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5173..5192
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5173..5191
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgaattggag ctccaccgcg gtggcggccg ctctagaact agtggatcca tgcagaacca
       61 gcagagcata tgggctcgct ggctgcacgt cgactctaga aaccagcaga gcatatgggc
      121 tcgctggatg cagaaccagc agagcatatg ggctcgctgg ctgcacgtcg actctagaaa
      181 ccagcagagc atatgggctc gctggatgca gaaccagcag agcatatggg ctcgctggct
      241 gcacgtcgac tctagaaacc agcagagcat atgggctcgc tggatgcaga accagcagag
      301 catatgggct cgctggctgc acgtcgactc tagaaaccag cagagcatat gggctcgctg
      361 gatgcagaac cagcagagca tatgggctcg ctggctgcac gtcgactcta gaaaccagca
      421 gagcatatgg gctcgctgga tgcagaacca gcagagcata tgggctcgct ggctgcacgt
      481 cgactctaga aaccagcaga gcatatgggc tcgctggatg catagatcta acccttaata
      541 taacttcgta taatgtatgc tatacgaagt tattaggtga tctgccactt ctaaataagc
      601 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
      661 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
      721 cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg accacacctc
      781 taccggcaga tccgctaggg ataacagggt ggatctgttt agcttgcctc gtccccgccg
      841 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
      901 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
      961 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
     1021 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
     1081 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
     1141 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
     1201 tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1261 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     1321 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1381 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1441 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1501 gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1561 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1621 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     1681 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     1741 ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     1801 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     1861 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     1921 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     1981 ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt
     2041 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     2101 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     2161 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     2221 tcgatactaa cgccgccatc cagtgtcgaa aacgagctct cgagaaccct taatataact
     2281 tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag tggcctatgc
     2341 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     2401 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2461 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2521 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2581 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2641 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2701 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2761 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2821 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2881 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2941 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3001 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3061 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3121 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3181 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3241 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3301 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3361 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3421 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3481 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3541 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3601 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3661 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3721 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3781 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3841 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3901 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3961 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4021 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4081 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4141 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4201 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4381 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4441 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4501 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     4561 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4621 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4681 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4741 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4801 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4861 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4921 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4981 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa
     5041 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     5101 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     5161 agtgagcgcg cgtaatacga ctcactatag gg
//
