LOCUS       sequence-434045-        8018 bp    DNA     circular SYN 14-APR-2025
DEFINITION  part A of YAP/SPARK-ON.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8018)
  AUTHORS   Chung CI, Yang J, Shu X
  TITLE     Chemogenetic Minitool for Dissecting the Roles of Protein Phase 
            Separation.
  JOURNAL   ACS Cent Sci. 2023 Jul 7;9(7):1466-1479. doi: 
            10.1021/acscentsci.3c00251. eCollection 2023 Jul 26.
  PUBMED    37521779
REFERENCE   2  (bases 1 to 8018)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acscentsci.3c00251"; journalName: "ACS Cent Sci"; date: 
            "2023-07-7- 7"; volume: "9"; issue: "7"; pages: "1466-1479"
COMMENT     Sequence Label: CEL-EGFP-YAP
FEATURES             Location/Qualifiers
     source          1..8018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1295..1297,1298..1300,1301..2011)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1340..1361)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1601..1620)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1948..1969
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3609..3626)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3615..3839
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3885..4313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3972..3991)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4182..4203
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4322..4342)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4327..4656
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4507..4642
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4569..4588
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4723..5517
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4777..4796)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5387..5406
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5691..5812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5728..5747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5782..5801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5861..5877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5861..5877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5874..5896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5885..5901
                     /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(5909..5915,5916..5933,5934..5939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5954..5975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6092..6109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6263..6851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6343..6362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7022..7813,7814..7882))
                     /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     7645..7664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7883..7987)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgtccct ttgctgtaaa caatgtcaag aaacagaaat
      961 aacaaccaaa aatgaaatat tcagtttatc cttatgtggg ccgatggcag cttatgtgaa
     1021 tcctcatgga tatgtgcatg agacacttac tgtgtataag gcttgcaact tgaatctgat
     1081 aggccggcct tctacagaac acagctggtt tcctgggtat gcctggactg ttgcccagtg
     1141 taagatctgt gcaagccata ttggatggaa gtttacggcc accaaaaaag acatgtcacc
     1201 tcaaaaattt tggggcttaa cgcgatctgc tctgttgccc acgatcccag acctcgagtc
     1261 cggactcaga tctggcagcg gtggaggcga tatcatggtg agcaagggcg aggagctgtt
     1321 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1381 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1441 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1501 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1561 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1621 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1681 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1741 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1801 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1861 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1921 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1981 gatcactctc ggcatggacg agctgtacaa gggatccggc agtgctggtg gtagtgctgg
     2041 tggtagtgct ggtggtagtg ctggtggcag tgctggtggt agtgctggtg gtagtgctgg
     2101 tggctctaga atggatcccg ggcagcagcc gccgcctcaa ccggcccccc agggccaagg
     2161 gcagccgcct tcgcagcccc cgcaggggca gggcccgccg tccggacccg ggcaaccggc
     2221 acccgcggcg acccaggcgg cgccgcaggc accccccgcc gggcatcaga tcgtgcacgt
     2281 ccgcggggac tcggagaccg acctggaggc gctcttcaac gccgtcatga accccaagac
     2341 ggccaacgtg ccccagaccg tgcccatgag gctccggaag ctgcccgacg ccttcttcaa
     2401 gccgccggag cccaaatccc actcccgaca ggccagtact gatgcaggca ctgcaggagc
     2461 cctgactcca cagcatgttc gagctcattc ctctccagct tctctgcagt tgggagctgt
     2521 ttctcctggg acactgaccc ccactggagt agtctctggc ccagcagcta cacccacagc
     2581 tcagcatctt cgacagtctt cttttgagat acctgatgat gtacctctgc cagcaggttg
     2641 ggagatggca aagacatctt ctggtcagag atacttctta aatcacatcg atcagacaac
     2701 aacatggcag gaccccagga aggccatgct gtcccagatg aacgtcacag cccccaccag
     2761 tccaccagtg cagcagaata tgatgaactc ggcttcaggt cctcttcctg atggatggga
     2821 acaagccatg actcaggatg gagaaattta ctatataaac cataagaaca agaccacctc
     2881 ttggctagac ccaaggcttg accctcgttt tgccatgaac cagagaatca gtcagagtgc
     2941 tccagtgaaa cagccaccac ccctggctcc ccagagccca cagggaggcg tcatgggtgg
     3001 cagcaactcc aaccagcagc aacagatgcg actgcagcaa ctgcagatgg agaaggagag
     3061 gctgcggctg aaacagcaag aactgcttcg gcaggagtta gccctgcgta gccagttacc
     3121 aacactggag caggatggtg ggactcaaaa tccagtgtct tctcccggga tgtctcagga
     3181 attgagaaca atgacgacca atagctcaga tcctttcctt aacagtggca cctatcactc
     3241 tcgagatgag agtacagaca gtggactaag catgagcagc tacagtgtcc ctcgaacccc
     3301 agatgacttc ctgaacagtg tggatgagat ggatacaggt gatactatca accaaagcac
     3361 cctgccctca cagcagaacc gtttcccaga ctaccttgaa gccattcctg ggacaaatgt
     3421 ggaccttgga acactggaag gagatggaat gaacatagaa ggagaggagc tgatgccaag
     3481 tctgcaggaa gctttgagtt ctgacatcct taatgacatg gagtctgttt tggctgccac
     3541 caagctagat aaagaaagct ttcttacatg gttataagaa ttcgggcccg tttaaacccg
     3601 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3661 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3721 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3781 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3841 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3901 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3961 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4021 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4081 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4141 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4201 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4261 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4321 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4381 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4441 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4501 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4561 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4621 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4681 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4741 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4801 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4861 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4921 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4981 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5041 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5101 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5161 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5221 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5281 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5341 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5401 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5461 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5521 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5581 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5641 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5701 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5761 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5821 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5881 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5941 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6001 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6061 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6961 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7021 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7141 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7621 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7981 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
