LOCUS       sequence-492079-        6677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert an N-terminal msGFP2 tag at VRG4 by popin-popout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6677)
  TITLE     Glick lab plasmids 2025
REFERENCE   2  (bases 1 to 6677)
  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"
COMMENT     Sequence Label: YIplac211-msGFP2-Vrg4
FEATURES             Location/Qualifiers
     source          1..6677
                     /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             720..1427
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(3175..3192)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3175..3191)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3184..3206)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3400..3419)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3519..3541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3659..4462)
                     /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(4463..4678)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4685..4703)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4771..4875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4876..4944,4945..5736)
                     /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(5094..5113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5907..6495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6396..6415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6649..6666
                     /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 ccaagcttgt
      241 aatggaccat ggcaatgtgc gccatatata tatgcggctc ccttatgagt agaagaaaat
      301 atatgactgg tggtacaaag tggcatggtg gtatatagat gtaggaacac catttgagtt
      361 tgccataatg attaaatttc cctttttata aagaattagc cgcccataac acgaaaaata
      421 tcacagttga cgaaagaaga cacgtcgcct aaacgcgtaa tgaatcgcaa tatacgaaaa
      481 gctaaacggg tttttcagaa tcctcttacc cagccttgga taatagtttt ggccgaaaaa
      541 aaaaaaaaga aagaaaaaaa ttgaaatttc gcgatccgaa caaacaatga acagaaaaac
      601 agaatatata taacgctgcc aagcctattt cttgttgccg attaattgtt gacatcctct
      661 taattaccaa aagagcctaa gaaaacaaac acactaacca cacagtatct ttcgcccgaa
      721 tggatagcac tgagagcttg ttcacaggtg ttgttcctat cttggtcgaa ttagacggtg
      781 acgtaaacgg tcataagttc agtgttagag gtgaaggtga aggtgacgct actaacggta
      841 aattgacatt gaagtttatt tgtactacag gtaaattgcc agttccttgg ccaactttgg
      901 ttactacatt gacatacggt gttcaatgtt tttcaagata cccagatcat atgaagcaac
      961 atgatttctt taagtctgct atgccagaag gttacgttca agaaagaact atcactttta
     1021 aagatgatgg tacttacaag acaagagcag aggttaagtt cgaaggtgac actttggtta
     1081 acagaatcga attgaagggt atcgatttta aagaagatgg taatattttg ggtcataaat
     1141 tggaatacaa ctacaactca cataacgttt acattacagc tgataaacaa aagaatggta
     1201 ttaaagcaaa cttcaagatc agacataatg ttgaagatgg ttctgttcaa ttagcagatc
     1261 attaccaaca aaacactcca attggtgacg gtccagtttt gttaccagat aaccattact
     1321 tgtctacaca atcaaaattg tctaaagatc caaacgaaaa gagagatcat atggttttgt
     1381 tagaatttgt tactgctgct ggtatcacag gtggcagtgg cggttcttca gaattgaaaa
     1441 caggtcatgc aggccataac ccttgggctt cagttgccaa ttccggtccg atctctattt
     1501 tatcctactg tggttcctct attttaatga cggtgactaa caagttcgtc gtcaatttga
     1561 aggatttcaa catgaacttt gtcatgcttt tcgtgcaatc tttggtttgt actataacct
     1621 tgattatcct acgtatactg ggctatgcga agttccgttc attaaacaaa acagacgcca
     1681 agaactggtt ccctatttcc tttttactgg tcttgatgat ctacacctct tcgaaggctt
     1741 tacaatactt ggctgttcca atttacacca ttttcaagaa tttgactatt atcttgattg
     1801 cttatggtga ggttctcttt tttggtggct ctgtcacctc catggaattg tcatcatttt
     1861 tgttgatggt cctttcttct gtcgttgcaa cttggggtga ccagcaagct gtggctgcca
     1921 aggctgcttc attggctgaa ggagcagccg gtgctgttgc ctcctttaac ccaggttatt
     1981 tctggatgtt caccaactgt atcacttctg cattattcgt tcttataatg agaaagagaa
     2041 ttaagttaac taacttcaag gatttcgaca ctatgtttta caacaatgtt ttggctctac
     2101 ctattctatt gctgttttct ttctgtgtgg aagattggtc ttcagttaat ttgaccaata
     2161 acttttctaa cgattcgcta actgctatga tcatcagtgg tgttgcatcc gtcggtattt
     2221 cttactgttc cggttggtgt gttcgtgtta cttcgtctac tacatattcg atggtagggg
     2281 ctttgaacaa gctgccaatt gccttgtctg gtttgatttt ctttgatgct ccaagaaact
     2341 tcttatctat tctctccatt tttattggtt tcctatcagg tattatttat gctgttgcca
     2401 aacaaaagaa gcaacaagcc caacctttac gtaaatgaga acttacgggg ggtgcaattt
     2461 attttttttt tttggtttat tttattttat agaggcatct aatgcaagta gatttatata
     2521 caattatact taaaattgat ataccttaga acaggtcgga cacaacgtgt ggtaactcca
     2581 atatatcact aatgacaata gcgccttctg gtgtttgccc caaaatttca tttacttcat
     2641 tctcaactaa atgaatacat gttttcatgc ccaatttgat acctgtctcg atgtttttcc
     2701 cactgtcatc tataaaataa gcattctcgt atcttgccaa cccgctttcc ttcatggctt
     2761 tttcaaatgc tttaacatgt ggtttgcaga ccagcgtatc tgtcctagag tagtcacaat
     2821 atgtcaatcc atcaaataaa tccgctatac ccaacaatct caggcaacgt attgcgtggt
     2881 ttttataagc gttggtgaaa agccacaatt tatcaatctt tcccgattgt ctcaatctca
     2941 acagcatatt ccttaatggt atatcaggtt tcaagatatc ttgtagtggt aaggaatcat
     3001 ccacaagccg gttatattcc agtgcgttca ctttgtggaa cattacaagt ccccttatgg
     3061 caagtccata ctctttgtag taagagttat tcaaaacgtg agcatcttca ggtgatagtt
     3121 tcaagtgtgt ttgaaagaac ctcagtatag attgttgcat aagatcgtga attcactggc
     3181 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     3241 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3301 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     3361 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3421 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3481 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3541 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc gggtaataac tgatataatt
     3601 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
     3661 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
     3721 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
     3781 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     3841 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     3901 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     3961 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
     4021 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
     4081 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     4141 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     4201 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     4261 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     4321 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     4381 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     4441 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcatgttttt gttctgtgca
     4501 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     4561 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     4621 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     4681 gctggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4741 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4801 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4861 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4921 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4981 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5041 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5101 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5161 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5221 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5281 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5341 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5401 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5461 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5521 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5581 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5641 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5701 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5761 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5821 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5881 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5941 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6001 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6061 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6121 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6181 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6241 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6301 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6361 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6421 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6481 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6541 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6601 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6661 agcgaggaag cggaaga
//
