LOCUS       sequence-402489-        5515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast vector encoding donor DNA for integration of 40nm GEMs into 
            the HO locus. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5515)
  AUTHORS   Shu T et al.
  TITLE     nucGEMs probe the biophysical properties of the nucleoplasm
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 5515)
  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-Pfv-Sapphire-HO
FEATURES             Location/Qualifiers
     source          1..5515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..87)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(241..829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(321..340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1012..1106)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             join(1124..1192,1193..1984)
                     /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(1342..1361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2155..2173
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2177..2220)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             4000..4716
                     /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      4737..4924
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4792..4816)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      join(5504..5515,1..46)
                     /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 gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat
       61 gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      241 tttccacagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgt
      781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      841 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      901 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      961 atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg gattctcacc
     1021 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     1081 cattactgga tctatcaaca ggagtccaag cgagctcgat atcatgagta ttcaacattt
     1141 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1201 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1261 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1321 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     1381 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1441 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1501 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1561 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1621 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     1681 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1741 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1801 ctggtttatt gctgataaat ctggagccgg tgagcgtgga tctcgcggta tcattgcagc
     1861 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1921 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1981 gtaatttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     2041 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc ccgatcaact
     2101 cgagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2161 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     2221 tggaattcgc ggccgcgtat ccttgcctaa cttcaatgtc aaagttgaaa tattctcctt
     2281 tagagcgctc catttcttct atgaagcgtt ttgcggcaaa ctcaccttca actgtcattg
     2341 ggaatgtctt atgatggttt tttggaatta ttattatcct accatcaagc gtctgacatt
     2401 gctgcagatt tctccatctc actttatatt tggtggcatt tctaccactt ttttccaaca
     2461 gtggtttggt agggaccctg actgacaatt tatgacctgc agtacattgt aatgcaagac
     2521 gctgataaac tgttctacgc ctgggatcta acctaccagg ttcaccttca aaagctctgt
     2581 gtttggtttt ttgctgtata ttatagattt tctgatagcc ctgtgtgaca tttatgacgc
     2641 gggcagcgga gccatctgcg cacataacgt aagagttagc cgtgacgttt gcgatgtctt
     2701 taatttcacc gttagcgaat tggagctctc ttggcttggt ttgtatgagc ttgttacgta
     2761 tcaaacaaaa tattcacatg tttttctcac cttaaagaca gaggcgcgat tgcgtttgcg
     2821 aaattccaat gccaacaaga taagcttgac gaaggagtta agagggcggc ttgaactaaa
     2881 aagagaaaag caatatctag aatgctctca ataaatccaa cccttataaa tagggataag
     2941 ccatacacca aagaagaatt aatggaaatt ctgagattag caataatagc tgagcttgat
     3001 gcaataaatc tatatgagca gatggcaaga tattccgaag atgaaaacgt aagaaagatt
     3061 ctccttgatg ttgcaaggga ggagaaggct cacgttggag aattcatggc actccttcta
     3121 aatttggatc cagaacaagt aactgagctg aagggtggat ttgaggaagt caaggagcta
     3181 actggaattg aggctcacat aaatgataac aagaaggagg aatctaacgt ggaatatttt
     3241 gagaagttga gaagtgctct tttggatggg gtaaataagg gaagatcttt actaaagcac
     3301 ttgccagtaa cgagaataga aggacaatca ttcagggttg acatcattaa atttgaagat
     3361 ggtgttaggg tagtaaagca agagtacaag ccaattccat tattgaagaa gaaattctat
     3421 gtgggaataa gagaactcaa cgatgggact tacgatgtat ctatagcaac taaagctggg
     3481 gaactcctag ttaaagacga agagtcgcta gtaataagag aaattctatc cacagaggga
     3541 attaagaaaa tgaaactttc ctcctgggac aatccagaag aagcacttaa tgatttaatg
     3601 aacgcactgc aggaagcctc aaatgcgtca gctggaccat ttgggctgat aattaaccca
     3661 aaaagatatg caaagttgct aaaaatttac gagaagagtg gaaagatgct agttgaagtg
     3721 ctaaaagaga tattcagggg aggaatcatt gttactctaa acattgatga gaataaggta
     3781 ataatatttg ccaatacacc agctgttctt gacgttgtag ttggccaaga tgtgactcta
     3841 caagagctag gacctgaagg agacgacgtt gcattcttgg ttagtgaagc tataggaata
     3901 agaataaaga acccagaggc aatagtagtc ttagagtggt ctggcagcgg tagtgggtcg
     3961 ggatcaggct ccggtcgacg gatccccggg ttaattaaga tgtctaaagg tgaagaatta
     4021 ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt
     4081 tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt
     4141 tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt ttcttatggt
     4201 gttcaatgtt ttgctagata cccagatcat atgaaacaac atgacttttt caagtctgcc
     4261 atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg taactacaag
     4321 accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt
     4381 attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa ctttaactct
     4441 cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt
     4501 agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca
     4561 attggtgatg gtccagtctt gttaccagac aaccattact tatccattca atctgcctta
     4621 tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct
     4681 ggtattaccc atggtattga tgaattgtac aaataaggcg cgccacttct aaataagcga
     4741 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     4801 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     4861 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
     4921 ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtcatgc
     4981 tttaaagtat agatagaatt gattgctgct tatgaggata tggatttaga gctaatagtt
     5041 gtaatagaaa taaaagccat ttagaatagg aattgaatac aattttatga gctttaaata
     5101 tttaatgatg aatagtgcgt ttagaacgct tcatcatgct tcaacaagct tctgcaaacc
     5161 tcatttgagt agcaatagct gtttactttt ttccgcgact ttattcgaag ttatggtgaa
     5221 cttttcgtgg tttcacacct aataacgccc agcggtagta caattcaaag tagtaggtac
     5281 caatggtagt actagtgggt gatagtagca aatttctcta tgaaaatgaa ttgttgctct
     5341 gcaatgatgt taatgtggac atgacatgaa ttttcgcgat atgttgatgt aaataataac
     5401 atgattttcg tggatcattt cgtaaaccat aggtttattt cgtgaattac taagagaaag
     5461 gcatagaaac aggacttgcg gatgggatac gcggccgctg cagtccggca aaaaa
//
