LOCUS       sequence-156222-        9992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated L-A capsid for curing satellite virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9992)
  TITLE     LACap
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9992)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pESC-NAT-LACap
FEATURES             Location/Qualifiers
     source          1..9992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2107..2296
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2368..2385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2539..3127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2619..2638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3298..4089,4090..4158))
                     /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     3921..3940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4159..4263)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4290..5632
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5253..5300)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     5673..5691
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5729..5751)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5851..5870
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            6517..7636
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        6517..6860
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6861..7433
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      7439..7636
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(8375..8830)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8512..8533)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8724..8743
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(8913..9078)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     8999..9023
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(9226..9249)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        9296..9960
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    9512..9629
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     9924..9947
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     9971..9990
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9971..9989
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cattcatata actccccatg ctaagatttg ttactaaaaa ctctcaagat aaatcgtctg
       61 atctattctc tatttgttct gatcgcggaa cttttgttgc ccataataga gttcggactg
      121 atttcaagtt tgacaactta gtattcaacc gagtttatgg tgtttctcaa aaatttacgt
      181 tagtcggtaa cccgacagtc tgctttaatg aaggcagttc ttacctagaa ggtattgcta
      241 aaaagtacct aactttggat ggaggacttg ccattgacaa tgtcctcaat gagttgagat
      301 ccacttgcgg tatcccaggt aatgctgttg catctcatgc gtataatatt acatcctggc
      361 gttggtatga caatcacgtg gcactgttga tgaacatgtt gcgtgcttac cacttacaag
      421 tattgaccga acagggccaa tatagcgctg gagatatccc tatgtaccat gatggacatg
      481 tcaaaatcaa gctaccagtg actatcgatg acacggcagg cccaacacaa ttcgcttggc
      541 ctagtgacag gtctactgat tcgtatcctg attgggcaca gttttctgaa tcatttccat
      601 caatcgacgt cccgtaccta gatgttaggc cattgaccgt aacggaagtc aatttcgtgc
      661 ttatgatgat gagtaagtgg catagacgta ctaacttagc gatagactac gaggcacccc
      721 aactagctga taagttcgct taccgccatg cgcttactgt tcaagacgct gacgagtgga
      781 tagaaggcga tagaactgat gaccagttcc gccccccctc gtctaaagta atgttatcgg
      841 cacttcgtaa gtacgtgaac cataacaggc tgtacaatca gttttacact gcagcacaac
      901 tgttagctca aattatgatg aaacctgtcc ctaactgcgc tgagggctac gcttggctga
      961 tgcatgacgc attggtcaat ataccaaaat ttgggtctat tcgaggaagg tacccctttt
     1021 tgttatcagg tgatgcagcg ttgattcagg ctacagccct agaagactgg tctgctatca
     1081 tggcgaaacc cgagctggtg ttcacttacg cgatgcaggt gtcagtagcg ttaaacaccg
     1141 gactatactt acgtcgcgtt aagaaaacag gcttcggcac aactatagat gacagctatg
     1201 aagatggagc gtttttgcaa ccggagacgt tcgttcaggc cgcactagca tgttgtaccg
     1261 gacaagatgc gcccctaaat gggatgtcag atgtgtatgt cacttatcca gatcttctag
     1321 aatttgatgc tgttacacaa gtacccatca cggtcattga gcccgctggc tataacattg
     1381 ttgatgatca tttagtggtt gtgggtgtac ctgtggcatg ttcaccatac atgatatttc
     1441 cagtagctgc gtttgatact gcaaatcctt actgtgggaa ttttgtcatt aaggctgcta
     1501 acaagtatct ccgtaagggt gccgtgtatg ataaactcga agcatggaag ttggcctggg
     1561 cactgagggt agccgggtat gacactcact tcaaagtgta tggcgataca cacggcttaa
     1621 ctaagttcta tgctgacaac ggtgacacat ggacacacat acctgaattt gtcactgacg
     1681 gtgacgtgat ggaagtattc gttactgcca tcgaacgcag agctagacat ttcgttgaac
     1741 tacctagact gaattcacca gcattcttca gatctgtaga agtcagcacc actatatatg
     1801 atactcatgt gcaggctggt gcgcatgcgg tgtatcatgc tagtcgaatc aatcttgatt
     1861 atgttaagcc tgtttcgacc ggcattcagg tgatcaatgc gggcgaactt aagaactact
     1921 ggggtagtgt gcgtcgtact cagcagggtt taggagtggt aggtcttacg atgccagctg
     1981 taatgcctac cggagaacct acagctggcg ctgcccacga agagttgata gaacaggcgg
     2041 acaatgtttt agtagagtaa acgtaatcga acccctcgag taagcttggt accgcggcta
     2101 gctaagatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     2161 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     2221 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     2281 tttgggacgc tcgaagatcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2341 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2701 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2941 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4261 gcgcacattt ccccgaaaag tgccacctga acgaagcatc tgtgcttcat tttgtagaac
     4321 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     4381 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt
     4441 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     4501 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     4561 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     4621 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     4681 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     4741 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     4801 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     4861 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     4921 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     4981 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     5041 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     5101 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     5161 tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc
     5221 ttcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct agagaatagg
     5281 aacttcggaa taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc
     5341 gagctgcgca catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata
     5401 tatatataca tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc
     5461 gtctatttat gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg
     5521 cggggtatcg tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct
     5581 caattggatt agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcatactaag
     5641 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5701 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5761 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5821 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5881 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
     5941 accattatgg gaaatgcttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
     6001 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
     6061 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
     6121 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
     6181 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
     6241 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
     6301 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
     6361 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtgcagctg
     6421 aagcttcgta cgctgcaggt cgacggatcc ccgggttaat taaggcgcgc cagatctgtt
     6481 tagcttgcct tgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     6541 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     6601 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     6661 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     6721 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     6781 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     6841 acatccgaac ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca
     6901 ccagtgtccc gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca
     6961 ccgtcttccg cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc
     7021 cgcccctgac caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg
     7081 acggcgaccc ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct
     7141 tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg
     7201 ccccggagca ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg
     7261 cccgcgagcg gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga
     7321 tccacgcgta ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg
     7381 gcaccgcctc ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac
     7441 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     7501 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     7561 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     7621 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag
     7681 ctcgaattca tcgatgatat cagatccact agtggcctag gtaagaaaag ttagaaagta
     7741 agacgattgc taaccaccta ttggaaaaaa caataggtcc ttaaataata ttgtcaactt
     7801 caagtattgt gatgcaagca tttagtcatg aacgcttctc tattctatat gaaaagccgg
     7861 ttccggcctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgtc
     7921 aggcgacctc tgaaattaac aaaaaatttc cagtcatcga atttgattct gtgcgatagc
     7981 gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg gttgctaaga gattcgaact
     8041 cttgcatctt acgatacctg agtattccca cagttaactg cggtcaagat atttcttgaa
     8101 tcaggcgcct tagaccgctc ggccaaacaa ccaattactt gttgagaaat agagtataat
     8161 tatcctataa atataacgtt tttgaacaca catgaacaag gaagtacagg acaattgatt
     8221 ttgaagagaa tgtggatttt gatgtaattg ttgggattcc atttttaata aggcaataat
     8281 attaggtatg tggatatact agaagttctc ctcgaccgtc gatatgcggt gtgaaatacc
     8341 gcacagatgc gtaaggagaa aataccgcat caggaaattg taaacgttaa tattttgtta
     8401 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     8461 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     8521 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     8581 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     8641 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     8701 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     8761 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     8821 cagggcgcgt cgcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     8881 cgggcctctt cgctattacg ccagctgaat tggagcgacc tcatgctata cctgagaaag
     8941 caacctgacc tacaggaaag agttactcaa gaataagaat tttcgtttta aaacctaaga
     9001 gtcactttaa aatttgtata cacttatttt ttttataact tatttaataa taaaaatcat
     9061 aaatcataag aaattcgctt atttagaagt gtcaacaacg tatctaccaa cgatttgacc
     9121 cttttccatc ttttcgtaaa tttctggcaa ggtagacaag ccgacaacct tgattggaga
     9181 cttgaccaaa cctctggcga agaattgtta attaagagct cagatcttat cgtcgtcatc
     9241 cttgtaatcc atcgatacta gtgcggccgc cctttagtga gggttgaatt cgaattttca
     9301 aaaattctta cttttttttt ggatggacgc aaagaagttt aataatcata ttacatggca
     9361 ttaccaccat atacatatcc atatacatat ccatatctaa tcttacttat atgttgtgga
     9421 aatgtaaaga gccccattat cttagcctaa aaaaaccttc tctttggaac tttcagtaat
     9481 acgcttaact gctcattgct atattgaagt acggattaga agccgccgag cgggtgacag
     9541 ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac
     9601 gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta
     9661 tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa atgaacgaat
     9721 caaattaaca accataggat gataatgcga ttagtttttt agccttattt ctggggtaat
     9781 taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa aaactgcata
     9841 accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt caaatgtaat
     9901 aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac
     9961 cccggatccg taatacgact cactataggg cc
//
