LOCUS       sequence-492071-        6425 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Insert a C-terminal moxGFP2 tag at KRE2 by popin-popout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6425)
  TITLE     Glick lab plasmids 2025
REFERENCE   2  (bases 1 to 6425)
  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-Kre2-moxGFP2
FEATURES             Location/Qualifiers
     source          1..6425
                     /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"
     primer_bind     complement(2923..2940)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2923..2939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2932..2954)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3148..3167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3267..3289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3407..4210)
                     /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(4211..4426)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4433..4451)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4519..4623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4624..4692,4693..5484)
                     /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(4842..4861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5655..6243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6144..6163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6397..6414
                     /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 ccaagcttgc
      241 atgcgccaaa agcttgttac gtcactttgg tgagaaacaa ggagttgaaa ggtttgctaa
      301 gctccattaa atatgtggaa aacaaaatta acaagaaatt cccatatcct tgggttttcc
      361 taaacgatga accttttact gaagaattca aggaagcagt caccaaagct gtttcttccg
      421 aagttaagtt tggtattttg cccaaggaac attggtcata tcctgaatgg attaatcaaa
      481 ccaaggctgc tgaaattcgt gcagatgctg ccaccaaata catatacggt ggctccgaat
      541 cttatagaca catgtgtcgt taccaatctg ggtttttctg gagacatgaa ttattagaag
      601 agtacgattg gtactggcgt gtggaaccag acatcaagtt atactgtgat attaattacg
      661 acgtttttaa gtggatgcaa gaaaacgaaa aagtttacgg ctttaccgtt tctattcatg
      721 aatatgaagt gacgatccca acactatggc aaacgtccat ggatttcatc aaaaagaacc
      781 ccgaatactt agatgaaaac aacctgatga gttttctttc gaacgataat ggtaaaacat
      841 acaatctgtg ccatttctgg tcaaactttg aaattgcaaa cttgaatttg tggaggtcac
      901 cagcctacag agagtatttt gacactttgg atcatcaagg tggatttttc tacgaaagat
      961 ggggcgatgc tcccgttcat tctattgctg ctgctttgtt tttgccaaag gataaaatcc
     1021 attatttttc agacattggt taccatcatc caccttatga taactgccca ttggacaagg
     1081 aggtctataa cagtaacaac tgtgaatgtg accaaggtaa tgatttcact ttccaaggtt
     1141 actcttgtgg taaggaatat tatgatgctc aagggttggt aaagccaaaa aactggaaaa
     1201 aattccgtga gggaagttcg atggatagca ctgagagctt gttcacaggt gttgttccta
     1261 tcttggtcga attagacggt gacgtaaacg gtcataagtt cagtgttaga ggtgaaggtg
     1321 aaggtgacgc tactaacggt aaattgacat tgaagtttat tagtactaca ggtaaattgc
     1381 cagttccttg gccaactttg gttactacat tgacatacgg tgttcaagtt ttttcaagat
     1441 acccagatca tatgaagcaa catgatttct ttaagtctgc tatgccagaa ggttacgttc
     1501 aagaaagaac tatcactttt aaagatgatg gtacttacaa gacaagagca gaggttaagt
     1561 tcgaaggtga cactttggtt aacagaatcg aattgaaggg tatcgatttt aaagaagatg
     1621 gtaatatttt gggtcataaa ttggaataca actacaactc acataacgtt tacattacag
     1681 ctgataaaca aaagaatggt attaaagcaa acttcaagat cagacataat gttgaagatg
     1741 gttctgttca attagcagat cattaccaac aaaacactcc aattggtgac ggtccagttt
     1801 tgttaccaga taaccattac ttgtctacac aatcaaaatt gtctaaagat ccaaacgaaa
     1861 agagagatca tatggttttg ttagaatttg ttactgctgc tggtatcaca taaaaatctt
     1921 ggaacatact gtttctttgt tttgacttta tactttctat ttatatttta tttttataac
     1981 tggttaagta cacataggac tgcgtatcaa acatataagt gaggcaatcc acattttttt
     2041 taaagattcg aatattttta ttctcattag cgtattccga gaatagttcg aaaaaatata
     2101 aggtatatca agagttttta caagtgagag gaaagaggaa taagctataa gcaacaaaag
     2161 cgtaaaaaaa ttagctgaag acatagaact atggatgttc tcaaagaggt gttgtcacta
     2221 gaccaagata aatttgacca gctgaaggaa acgagccgag ataaaacaaa tgaaacggat
     2281 gatccttttg aaaactattt gaaggattgt aaatttaaag cgccttcaaa caaagatcag
     2341 tcaccatttg ctaaacttaa atcattacag gaaactcatt ctaacaatga agcggctatt
     2401 aatataatta ttcctcaatt gattgattac ttaaccgaat tcactaatag gttatcaaat
     2461 tacacacaag atttagactt cattaaaaaa aagtccaatg aattacagtc attgctcgaa
     2521 tacaactcca ctaaactggc acatatctct cctatggtta atgatttgat gattcctcct
     2581 gaactcattg atgacatcat taaagggaag atcaatgaaa gctggcagga taatataaca
     2641 ttcatagcag ataaagaaga aatttataac aagtataggt ccaataatct cgatcaagac
     2701 aacaaggacg cagaaaattc agcaatgcta gcaccaaagg attttgataa gttatgtcaa
     2761 ctcctggaca tcctaaaaaa tgttattcta gaaaggtcga aaagacttat tatttccaaa
     2821 atcaaaactt tgaggagtca taacccagta ccatcgcaaa ggatacaaaa caaattatta
     2881 aaagttcaaa aaattttccc cttcggtacc gagctcgaat tcactggccg tcgttttaca
     2941 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3001 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3061 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3121 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3301 atcaccgaaa cgcgcgagac gaaagggcgg gtaataactg atataattaa attgaagctc
     3361 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     3421 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     3481 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     3541 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     3601 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     3661 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     3721 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     3781 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     3841 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     3901 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     3961 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     4021 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     4081 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     4141 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     4201 ctttcgacat gatttatctt cgtttcctgc atgtttttgt tctgtgcagt tgggttaaga
     4261 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     4321 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     4381 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagc tggcctcgtg
     4441 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4501 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4561 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4621 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4681 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4741 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4801 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4861 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4921 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4981 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5041 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5101 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5161 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5221 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5281 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5341 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5401 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5461 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5521 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgccagaccc cgtagaaaag
     5641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5761 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6301 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6361 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6421 gaaga
//
