LOCUS       sequence-492080-        8232 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Insert a C-terminal msGFP2x4 tag at RIC1 by popin-popout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8232)
  TITLE     Glick lab plasmids 2025
REFERENCE   2  (bases 1 to 8232)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: YIplac211-Ric1-msGFP2x4
FEATURES             Location/Qualifiers
     source          1..8232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(1107..1814)
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     CDS             complement(1821..2528)
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     CDS             complement(2535..3242)
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     CDS             complement(3249..3956)
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(4730..4747)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4730..4746)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4739..4761)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4955..4974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5074..5096
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5214..6017)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6018..6233)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6240..6258)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6326..6430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6431..6499,6500..7291)
                     /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(6649..6668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7462..8050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7951..7970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8204..8221
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccattgcctt
      241 gttcttttca gtgcagtggc agatgttacg agggtcaaat aatttttgaa tatatataga
      301 aatggctact tactataatt aaaaataaat atgaaaaata aaacttcagt accccacatc
      361 attctcattt ttttctgttt gtttgatttt taggaatttg gctccatcat gaacattcgt
      421 tagcgtatga tatcactttt tcattatctc ctattgttac tttacttttc atatagatat
      481 ccttcatccc gtgtaggtct actacgcaaa aatgaattct agtaattaat aataatgata
      541 taaaaaaggc taagtgcaca ttactaaaaa cgaaactaag atccttagtc tgagttgata
      601 tcacctgcga aaataccttt ttctctttga tcatcccatt tttcgatcca gtctagggga
      661 cataaggcgt tataggtctt ccaaaagacc ttgcacggag caaaatcttc gcccttcata
      721 ttaacacact tgtgataatc cacataagat tgccaacaat gctttgtttg gttttgttgg
      781 ggaaatctag catcgaaacc aactgtatgt agtggagagt tttcttgatc agccatgtct
      841 attatactgt atttatgtgc ttgttcctat ttgtgaaagc aaagatactt tatgtattct
      901 tactaaaata tttgtatatc tctgctaatc attttaatgc aacttttcta tctggtgcat
      961 ctttcatcac cttttaaaga tcctgaaggg atggggatat cgggcaaaac tacgcgcgac
     1021 gcatttctga aaattgattg gtcaatgtta tgaagctata tggtaaagaa tacatggtgt
     1081 catatagatg cggccgctta gctagcagaa ccgccactgc cacctgtgat accagcagca
     1141 gtaacaaatt ctaacaaaac catatgatct ctcttttcgt ttggatcttt agacaatttt
     1201 gattgtgtag acaagtaatg gttatctggt aacaaaactg gaccgtcacc aattggagtg
     1261 ttttgttggt aatgatctgc taattgaaca gaaccatctt caacattatg tctgatcttg
     1321 aagtttgctt taataccatt cttttgttta tcagctgtaa tgtaaacgtt atgtgagttg
     1381 tagttgtatt ccaatttatg acccaaaata ttaccatctt ctttaaaatc gatacccttc
     1441 aattcgattc tgttaaccaa agtgtcacct tcgaacttaa cctctgctct tgtcttgtaa
     1501 gtaccatcat ctttaaaagt gatagttctt tcttgaacgt aaccttctgg catagcagac
     1561 ttaaagaaat catgttgctt catatgatct gggtatcttg aaaaacattg aacaccgtat
     1621 gtcaatgtag taaccaaagt tggccaagga actggcaatt tacctgtagt acaaataaac
     1681 ttcaatgtca atttaccgtt agtagcgtca ccttcacctt cacctctaac actgaactta
     1741 tgaccgttta cgtcaccgtc taattcgacc aagataggaa caacacctgt gaacaagctc
     1801 tcagtgctat ccatactagc agaaccgcca ctgccacctg tgataccagc agcagtaaca
     1861 aattctaaca aaaccatatg atctctcttt tcgtttggat ctttagacaa ttttgattgt
     1921 gtagacaagt aatggttatc tggtaacaaa actggaccgt caccaattgg agtgttttgt
     1981 tggtaatgat ctgctaattg aacagaacca tcttcaacat tatgtctgat cttgaagttt
     2041 gctttaatac cattcttttg tttatcagct gtaatgtaaa cgttatgtga gttgtagttg
     2101 tattccaatt tatgacccaa aatattacca tcttctttaa aatcgatacc cttcaattcg
     2161 attctgttaa ccaaagtgtc accttcgaac ttaacctctg ctcttgtctt gtaagtacca
     2221 tcatctttaa aagtgatagt tctttcttga acgtaacctt ctggcatagc agacttaaag
     2281 aaatcatgtt gcttcatatg atctgggtat cttgaaaaac attgaacacc gtatgtcaat
     2341 gtagtaacca aagttggcca aggaactggc aatttacctg tagtacaaat aaacttcaat
     2401 gtcaatttac cgttagtagc gtcaccttca ccttcacctc taacactgaa cttatgaccg
     2461 tttacgtcac cgtctaattc gaccaagata ggaacaacac ctgtgaacaa gctctcagtg
     2521 ctatccatac tagcagaacc gccactgcca cctgtgatac cagcagcagt aacaaattct
     2581 aacaaaacca tatgatctct cttttcgttt ggatctttag acaattttga ttgtgtagac
     2641 aagtaatggt tatctggtaa caaaactgga ccgtcaccaa ttggagtgtt ttgttggtaa
     2701 tgatctgcta attgaacaga accatcttca acattatgtc tgatcttgaa gtttgcttta
     2761 ataccattct tttgtttatc agctgtaatg taaacgttat gtgagttgta gttgtattcc
     2821 aatttatgac ccaaaatatt accatcttct ttaaaatcga tacccttcaa ttcgattctg
     2881 ttaaccaaag tgtcaccttc gaacttaacc tctgctcttg tcttgtaagt accatcatct
     2941 ttaaaagtga tagttctttc ttgaacgtaa ccttctggca tagcagactt aaagaaatca
     3001 tgttgcttca tatgatctgg gtatcttgaa aaacattgaa caccgtatgt caatgtagta
     3061 accaaagttg gccaaggaac tggcaattta cctgtagtac aaataaactt caatgtcaat
     3121 ttaccgttag tagcgtcacc ttcaccttca cctctaacac tgaacttatg accgtttacg
     3181 tcaccgtcta attcgaccaa gataggaaca acacctgtga acaagctctc agtgctatcc
     3241 atactagcag aaccgccact gccacctgtg ataccagcag cagtaacaaa ttctaacaaa
     3301 accatatgat ctctcttttc gtttggatct ttagacaatt ttgattgtgt agacaagtaa
     3361 tggttatctg gtaacaaaac tggaccgtca ccaattggag tgttttgttg gtaatgatct
     3421 gctaattgaa cagaaccatc ttcaacatta tgtctgatct tgaagtttgc tttaatacca
     3481 ttcttttgtt tatcagctgt aatgtaaacg ttatgtgagt tgtagttgta ttccaattta
     3541 tgacccaaaa tattaccatc ttctttaaaa tcgataccct tcaattcgat tctgttaacc
     3601 aaagtgtcac cttcgaactt aacctctgct cttgtcttgt aagtaccatc atctttaaaa
     3661 gtgatagttc tttcttgaac gtaaccttct ggcatagcag acttaaagaa atcatgttgc
     3721 ttcatatgat ctgggtatct tgaaaaacat tgaacaccgt atgtcaatgt agtaaccaaa
     3781 gttggccaag gaactggcaa tttacctgta gtacaaataa acttcaatgt caatttaccg
     3841 ttagtagcgt caccttcacc ttcacctcta acactgaact tatgaccgtt tacgtcaccg
     3901 tctaattcga ccaagatagg aacaacacct gtgaacaagc tctcagtgct atccatacta
     3961 gaacctccac cggagccgcc tccagaacca ccacctggat ccttgaatct ctcgagtgct
     4021 ttttgaacca attgtgtgtt gttttctctg tctaattcct tgagtaagcg tattaattgg
     4081 aaacacatgt cccacgaatc ggtagcttta attgggtctg tggcgtttgc agcacttgtc
     4141 accaaaagct ctaagacttt aagcattaat tcttcatcct tcaataaatt agtgacagat
     4201 ttctctgact tttttgtaga gttatcattt agagctttgc tttcctcagt ccctaaatct
     4261 gatttcttaa aatgtagatc atctcccaaa tctttctcat cgtaatttaa aaacactaac
     4321 agaagtacgc caagcatttt agcctcattt tcttcgatac agagtgctaa taaatctctt
     4381 ggtgtcatct ttagattagt gaatagatgg ttccagtatt tagtttcgat cttccttaaa
     4441 caattactaa taatttctaa cataccactg tgggttggtg gtgaaatatt atgctcagga
     4501 tcggtgttgt catacatctt aattaatttc aggatatcat ccaaagagtc gcttcgcagt
     4561 attttagtag acaaaatctt ttcgagtgca aatttgtaag gctttaaaaa acgataattg
     4621 tgtgaaatat cctccaaatc tgtgttgtct tctaatttgg ctaagattaa ttggtcgagg
     4681 tatatttggt gtttcaccac caatttggaa aatcgcgcgt ctcgaattca ctggccgtcg
     4741 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4801 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4861 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     4921 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4981 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5041 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5101 caccgtcatc accgaaacgc gcgagacgaa agggcgggta ataactgata taattaaatt
     5161 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     5221 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     5281 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     5341 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     5401 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     5461 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     5521 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     5581 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     5641 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     5701 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     5761 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     5821 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     5881 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     5941 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     6001 tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct gtgcagttgg
     6061 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     6121 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     6181 tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaagctgg
     6241 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6301 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6361 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6421 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6481 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6541 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6601 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6661 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6721 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6781 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6841 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6901 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6961 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7021 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7081 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7141 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7201 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7261 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7321 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7381 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7441 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7501 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7561 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7621 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7681 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7741 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7801 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7861 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7921 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7981 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8041 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8101 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     8161 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     8221 ggaagcggaa ga
//
