LOCUS       sequence-402488-        5536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast vector encoding donor DNA for integration of nuclear 40nm GEMs
            into the HO locus. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5536)
  AUTHORS   Shu T et al.
  TITLE     nucGEMs probe the biophysical properties of the nucleoplasm
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 5536)
  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: "bioRxiv 
            2022"
COMMENT     Sequence Label: pINO4-NLS-Pfv-Sapphire-HO
FEATURES             Location/Qualifiers
     source          1..5536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..75)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(229..817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(309..328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1000..1094)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             join(1112..1180,1181..1972)
                     /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(1330..1349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2143..2161
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2165..2208)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             2893..2913
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4009..4725
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      4746..4933
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4801..4825)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      join(5513..5536,1..34)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
ORIGIN
        1 accaccctgc cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc
       61 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
      121 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct
      241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtgttt tttttgtttg
      781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      961 tatatatgag taaacttggt ctgacagctc gaggcttgga ttctcaccaa taaaaaacgc
     1021 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     1081 tatcaacagg agtccaagcg agctcgatat catgagtatt caacatttcc gtgtcgccct
     1141 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1201 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1261 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1321 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1381 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1441 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1501 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1561 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1621 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1681 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1741 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1801 tgataaatct ggagccggtg agcgtggatc tcgcggtatc attgcagcac tggggccaga
     1861 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1921 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aatttctcca
     1981 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     2041 ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaactcg agtgccacct
     2101 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2161 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg
     2221 ccgcgtatcc ttgcctaact tcaatgtcaa agttgaaata ttctccttta gagcgctcca
     2281 tttcttctat gaagcgtttt gcggcaaact caccttcaac tgtcattggg aatgtcttat
     2341 gatggttttt tggaattatt attatcctac catcaagcgt ctgacattgc tgcagatttc
     2401 tccatctcac tttatatttg gtggcatttc taccactttt ttccaacagt ggtttggtag
     2461 ggaccctgac tgacaattta tgacctgcag tacattgtaa tgcaagacgc tgataaactg
     2521 ttctacgcct gggatctaac ctaccaggtt caccttcaaa agctctgtgt ttggtttttt
     2581 gctgtatatt atagattttc tgatagccct gtgtgacatt tatgacgcgg gcagcggagc
     2641 catctgcgca cataacgtaa gagttagccg tgacgtttgc gatgtcttta atttcaccgt
     2701 tagcgaattg gagctctctt ggcttggttt gtatgagctt gttacgtatc aaacaaaata
     2761 ttcacatgtt tttctcacct taaagacaga ggcgcgattg cgtttgcgaa attccaatgc
     2821 caacaagata agcttgacga aggagttaag agggcggctt gaactaaaaa gagaaaagca
     2881 atatctagaa tgccaaaaaa gaagagaaag gtcctctcaa taaatccaac ccttataaat
     2941 agggataagc catacaccaa agaagaatta atggaaattc tgagattagc aataatagct
     3001 gagcttgatg caataaatct atatgagcag atggcaagat attccgaaga tgaaaacgta
     3061 agaaagattc tccttgatgt tgcaagggag gagaaggctc acgttggaga attcatggca
     3121 ctccttctaa atttggatcc agaacaagta actgagctga agggtggatt tgaggaagtc
     3181 aaggagctaa ctggaattga ggctcacata aatgataaca agaaggagga atctaacgtg
     3241 gaatattttg agaagttgag aagtgctctt ttggatgggg taaataaggg aagatcttta
     3301 ctaaagcact tgccagtaac gagaatagaa ggacaatcat tcagggttga catcattaaa
     3361 tttgaagatg gtgttagggt agtaaagcaa gagtacaagc caattccatt attgaagaag
     3421 aaattctatg tgggaataag agaactcaac gatgggactt acgatgtatc tatagcaact
     3481 aaagctgggg aactcctagt taaagacgaa gagtcgctag taataagaga aattctatcc
     3541 acagagggaa ttaagaaaat gaaactttcc tcctgggaca atccagaaga agcacttaat
     3601 gatttaatga acgcactgca ggaagcctca aatgcgtcag ctggaccatt tgggctgata
     3661 attaacccaa aaagatatgc aaagttgcta aaaatttacg agaagagtgg aaagatgcta
     3721 gttgaagtgc taaaagagat attcagggga ggaatcattg ttactctaaa cattgatgag
     3781 aataaggtaa taatatttgc caatacacca gctgttcttg acgttgtagt tggccaagat
     3841 gtgactctac aagagctagg acctgaagga gacgacgttg cattcttggt tagtgaagct
     3901 ataggaataa gaataaagaa cccagaggca atagtagtct tagagtggtc tggcagcggt
     3961 agtgggtcgg gatcaggctc cggtcgacgg atccccgggt taattaagat gtctaaaggt
     4021 gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt
     4081 cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta
     4141 aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt cactactttt
     4201 tcttatggtg ttcaatgttt tgctagatac ccagatcata tgaaacaaca tgactttttc
     4261 aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt
     4321 aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa
     4381 ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac
     4441 tttaactctc acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac
     4501 ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa
     4561 aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt atccattcaa
     4621 tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt
     4681 actgctgctg gtattaccca tggtattgat gaattgtaca aataaggcgc gccacttcta
     4741 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     4801 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     4861 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
     4921 acacctctac cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc
     4981 tcgtcatgct ttaaagtata gatagaattg attgctgctt atgaggatat ggatttagag
     5041 ctaatagttg taatagaaat aaaagccatt tagaatagga attgaataca attttatgag
     5101 ctttaaatat ttaatgatga atagtgcgtt tagaacgctt catcatgctt caacaagctt
     5161 ctgcaaacct catttgagta gcaatagctg tttacttttt tccgcgactt tattcgaagt
     5221 tatggtgaac ttttcgtggt ttcacaccta ataacgccca gcggtagtac aattcaaagt
     5281 agtaggtacc aatggtagta ctagtgggtg atagtagcaa atttctctat gaaaatgaat
     5341 tgttgctctg caatgatgtt aatgtggaca tgacatgaat tttcgcgata tgttgatgta
     5401 aataataaca tgattttcgt ggatcatttc gtaaaccata ggtttatttc gtgaattact
     5461 aagagaaagg catagaaaca ggacttgcgg atgggatacg cggccgctgc agtccggcaa
     5521 aaaagggcaa ggtgtc
//
