LOCUS       sequence-265756-        7117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of NAB2-mTagBFP2 as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7117)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 7117)
  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"; journalName: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM119
FEATURES             Location/Qualifiers
     source          1..7117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..64
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(102..124)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     224..243
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        324..342
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     324..341
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      3733..3920
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3788..3812)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3972..4005
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5008..5041
                     /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(5094..5113)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5095..5113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5200..5217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5371..5959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5451..5470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6130..6921,6922..6990))
                     /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     6753..6772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6991..7095)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
       61 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      121 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      181 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      241 gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata
      301 accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc gccagctgaa
      361 gcttcgtacg ctgcaggtcg tgggattcct gggttaatta acatcgctag cactaaaatc
      421 atagcgccct gtatcaaaaa gaagcttttt tcttttgttg ggtatgtcta tatagagcac
      481 atcctatatt cccttcgagg tgtttttaca agaacaaata aagaaacaaa ccgcggtgtg
      541 aggaagttcc cttaaaaatg gcaaagatta ccgtaaaggt aaattactcc tttaacgatg
      601 cttaaaacga gcatcttact cttgcaacga aattaatcac tcaatttcaa cgaggatgtt
      661 ggcttcccat caaagtagtt ttggggtaaa ccgcatatat ctgtatatta agaaaactta
      721 ttttccacta tctaatttga actacagaga agataagaaa tcttttaagt agcaatcatg
      781 ttttatctaa gtgacatcga ggaagaagcc tcggctgggg ctgaaccaac ttataatttc
      841 tgggaagttt tgttattctc caatactcaa gaaaacctcg tcaccgttgt cggagagttg
      901 cacacactta ccgatcgtgt ggttcattac aaaattgagc ctgaatcaag ggaagttact
      961 gccacaacac tgccatcttt gttagcactt ttgctagaaa aaaggaacca ggcgagacgt
     1021 ttatataggg atgtgctttc aatgaaaatg tcagaactgg actgggacat cgacgatttg
     1081 ttcacacagt tacaagaaga gctaacaaga actgatgaca ctctttctat gtatcctagg
     1141 agaaggttct atcactagct tgctcaacgc gaccttatac acagagttta atgtgaagag
     1201 aacattcata tacaattata taaacattgt tgtattctcg tacatcgtta cacaaaatac
     1261 gaagcgggga aatagtgata ttcgcttcgc tggtggcaaa tgggaaaaat acaaaatttt
     1321 acacatcttt cttccaaata cttccataaa acatctaaat atagtggttt ctaattatct
     1381 ctgatcataa ggaagtggaa gtacatcaga aatgtctcaa gaacagtaca cagaaaactt
     1441 gaaggttatc gttgccgaaa aactggctgg tataccaaac tttaacgaag atatcaagta
     1501 tgttgcggag tatattgtct tattgatcgt taacggtggt actgttgaat ctgtcgtaga
     1561 cgagctggct agtttgtttg atagtgtttc gagagatacg cttgcaaatg ttgttcaaac
     1621 agcctttttc gcattagaag ctctgcaaca gggagaaagt gctgaaaata ttgtttccaa
     1681 aattagaatg atgaatgcgc aaagcttggg acaatcggat atcgcacaac agcaacaaca
     1741 gcaacaacaa caacagcaac cagacatcgc gcaacagcaa cctcaacagc aacctcaaca
     1801 gcaacctcaa cagcaacctc aacagcaacc tcaacagcaa cctcaacagc aacctcaaca
     1861 gcaacctcaa cagcaacctc aacttcaacc acttcagcca caactaggga cccagaatgc
     1921 aatgcagaca gatgctcctg caactccatc ccccatatca gccttttccg gcgttgttaa
     1981 cgctgcagct ccccctcagt ttgcgcctgt agataacagc caaaggttca ctcaacgtgg
     2041 cggaggcgcc gttggaaaga atcgtagagg tggtcgcggt gggaaccgtg gaggacgcaa
     2101 caataattcc acacgtttta atccgttagc aaaagcactt ggaatggcgg gtgagagtaa
     2161 tatgaacttc actccaacca agaaagaggg gcgttgcaga ttgtttcctc actgtcctct
     2221 tggtagatca tgcccacatg cacacccaac taaggtatgt aatgaatatc caaattgtcc
     2281 aaagcctccc ggaacttgtg agtttttaca tccaaatgaa gatgaagagt tgatgaagga
     2341 aatggaaaga actcgtgaag aatttcaaaa aagaaaagct gatttattgg cggcaaaaag
     2401 gaaaccggta caaactggta tcgttctgtg taaatttggg gctctgtgtt ccaatccatc
     2461 atgcccattt ggtcatccaa caccagcaaa tgaagatgcg aaagtcattg atctaatgtg
     2521 gtgtgacaag aatttgacat gtgataatcc tgagtgtaga aaggcccact cttcattgtc
     2581 gaagatcaag gaagtaaaac caataagcca gaagaaagca gctccacctc cggttgaaaa
     2641 gtccttagaa caatgtaagt tcggtacgca ctgcaccaat aaacgttgca aatatagaca
     2701 tgctcgttct catattatgt gccgtgaagg agcaaactgt actagaattg attgtttatt
     2761 tggccatcca attaatgaag attgtagatt tggtgtcaat tgtaagaata tttactgtct
     2821 attcagacat cctccaggca gagtacttcc ggaaaagaaa ggcgctgcac ccaattcaaa
     2881 cgttcctacc aatgaaaggc catttgcatt gccagaaaac gcaataattg aaaatgctcc
     2941 tccgcaaacc agttttacgc accaagaaca agatacggaa atgaaccttt ctcttgaagt
     3001 tgctgaagaa atgagcgaac tgatcaaaga gaacatgcac atgaagctgt acatggaagg
     3061 caccgttgac aaccaccact ttaagtgcac ctctgagggt gagggtaagc cctacgaagg
     3121 cacccaaacc atgcgtatca aagttgtgga gggcggtcca ctgccattcg cttttgacat
     3181 tctggccacc agcttcctgt acggttccaa aaccttcatt aaccatactc agggcattcc
     3241 tgatttcttc aaacagagct ttccggaagg tttcacctgg gagcgtgtca ccacgtatga
     3301 agatggtggt gtgttgaccg ccacccaaga tacctccctg caagatggct gtctgatcta
     3361 taacgtgaaa attcgtggcg tcaactttac tagcaatggt cccgtgatgc agaagaaaac
     3421 cctgggttgg gaggccttta cagaaaccct gtatcctgcc gatggtggcc tggagggccg
     3481 taacgacatg gcactgaagc tggttggtgg cagccatttg atcgcaaatg ccaagaccac
     3541 ttacagaagc aagaaacccg caaaaaatct gaagatgccg ggtgtttact atgtcgacta
     3601 ccgtctggaa agaattaaag aagcaaataa tgagacttac gtggagcagc acgaggttgc
     3661 agtcgcgaga tattgcgact tgcctagcaa gctgggtcat aaactgaatt gaggcgcgcc
     3721 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     3781 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     3841 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt
     3901 attgaccaca cctctaccgg cagatccgct agggataaca gggtaatatc tgcaggtcga
     3961 caacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt ctagagcggc
     4021 cgcggaacga tcattcacta tatatatatc aatttatata tacgtatgtg taattgaaga
     4081 aagatacgtt ttttcctcta ttgagaggcc tgctggatga atagctttac cttttctaaa
     4141 tccttgatac catcagtctc tactcctcca cttacatcga caccgatcgc atttggtaac
     4201 atattaatgg caacagaaac gttatcagga ttcaatccac cagcgataat gaattttatc
     4261 tcgggatgac ttgcagacca actggaaatt gcactccaat tcaatttctc accagtgcca
     4321 ccttcaccag aatcgaacaa cgtcagcaca ttgtctacgt gttcatacag gtccagtagt
     4381 aattcacaat cctgtgggaa ctggaacctc ttaatgattg gaattgaaga tgggatcaaa
     4441 gatctgtatt ctttaatatc ttcatctcca tgtaattgta tcacatctag attatattcg
     4501 tggtacagtt gaaggacatc atcaacggac tgatttctaa acaccccgac tagtttagta
     4561 cctttcacgt tctcttgttg gtgaactgca gttgaaatac ctttcgcaac agatgagtca
     4621 atggttcttt tcctaccggg aacacaaatg atacctaagt aatcagcacc atcatccaca
     4681 gcagtctttg cagcttcaac ggtttgcaaa ccacacactt taacgagcat cacagtagat
     4741 ttcgcaagag ggtagctcgc tcagagtacc caagtaaatg attagtaaac tgatgtttga
     4801 tagttcaatt tttcaatgaa ataaccttat attaaaattg atattactat tatacaaaaa
     4861 taaagaataa aggatttgag tttatacata aaataccatt attatttgtt cagtgagaga
     4921 tactggggta tatgggatgt gtgtagtgat accatgcaat catgtatcaa acatgggccc
     4981 ggtaccgagc tctcgagaac ccttaatata acttcgtata atgtatgcta tacgaagtta
     5041 ttaggtgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata
     5101 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc
     5161 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5221 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5281 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5881 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5941 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6001 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6061 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6121 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6181 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6241 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6301 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6361 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6421 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6481 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6541 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6601 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6661 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6721 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6781 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6841 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6901 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6961 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7021 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7081 aataggggtt ccgcgcacat ttccccgaaa agtgcca
//
