LOCUS       sequence-370797-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the SkGAL2 promoter inserted with 2 PZ4 element using a 
            green fluorescent protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  AUTHORS   Peng B, Bandari NC, Lu Z, Howard CB, Scott C, Trau M, Dumsday G, 
            Vickers CE
  TITLE     Engineering eukaryote-like regulatory circuits to expand artificial 
            control mechanisms for metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Commun Biol. 2022 Feb 16;5(1):135. doi: 10.1038/s42003-022-03070-z.
  PUBMED    35173283
REFERENCE   2  (bases 1 to 5567)
  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: "Commun 
            Biol."; date: "2022-02-16"; volume: "5(1):135. doi"; pages: " 
            10.1038/s42003-022-03070-z"
COMMENT     Sequence Label: pILGFP10CF72 (B9)
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             29..739
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(984..1000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(984..1000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(997..1019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1008..1024
                     /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(1032..1038,1039..1056,1057..1062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1077..1098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1215..1232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1386..1974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1466..1485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2145..2936,2937..3005))
                     /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     2768..2787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3006..3110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3178..3196
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3234..3256)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3356..3375
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3569..3591
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3583..3600
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3584..3600
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4714..4747)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ataaaaaaag ttttttacaa tgggatcctc taaaggtgaa gaattattca ctggtgttgt
       61 cccaattttg gttgaattag atggtgatgt taatggtcac aaattttctg tctccggtga
      121 aggtgaaggt gatgctactt acggtaaatt gaccttaaaa tttatttgta ctactggtaa
      181 attgccagtt ccatggccaa ccttagtcac tactttcggt tatggtgttc aatgttttgc
      241 gagataccca gatcatatga aacaacatga ctttttcaag tctgccatgc cagaaggtta
      301 tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca gagctgaagt
      361 caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg attttaaaga
      421 agatggtaac attttaggtc acaaattgga atacaactat aactctcaca atgtttacat
      481 catggctgac aaacaaaaga atggtatcaa agttaacttc aaaattagac acaacattga
      541 agatggttct gttcaattag ctgaccatta tcaacaaaat actccaattg gtgatggtcc
      601 agtcttgtta ccagacaacc attacttatc cactcaatct gccttatcca aagatccaaa
      661 cgaaaagaga gaccacatgg tcttgttaga atttgttact gctgctggta ttacccatgg
      721 tatggatgaa ttgtacaaaa gatcttaaac tagtaggact atttgcaaag ggaagggatg
      781 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
      841 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
      901 gagcttcaat ttaattatat cagttattac ccgggaatct cggtcgtaat gatttaaatg
      961 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1021 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1081 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1141 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1201 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2221 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2281 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2341 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2401 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2461 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2521 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2581 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2641 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2701 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2761 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2821 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2881 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2941 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3001 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3061 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3121 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3181 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     3241 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3301 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     3361 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     3421 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     3481 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     3541 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     3601 gaattcattt aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     3661 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     3721 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaaacatgt
     3781 ccacaaaatc atataccagt agagctgaga ctcatgcaag tccggttgca tcgaaacttt
     3841 tacgtttaat ggatgaaaag aagaccaatt tgtgtgcttc tcttgacgtt cgttcgactg
     3901 atgagctatt gaaacttgtt gaaacgttgg gtccatacat ttgccttttg aaaacacacg
     3961 ttgatatctt ggatgatttc agttatgagg gtactgtcgt tccattgaaa gcattggcag
     4021 agaaatacaa gttcttgata tttgaggaca gaaaattcgc cgatatcggt aacacagtca
     4081 aattacaata tacatcgggc gtttaccgta tcgcagaatg gtctgatatc accaacgccc
     4141 acggggttac tggtgctggt attgttgctg gcttgaaaca aggtgcgcaa gaggtcacca
     4201 aagaaccaag gggattattg atgcttgctg aattgtcttc caagggttct ctagcacacg
     4261 gtgaatatac taagggtacc gttgatattg caaagagtga taaagatttc gttattgggt
     4321 tcattgctca gaacgatatg ggaggaagag aagaagggtt tgattggcta atcatgaccc
     4381 caggtgtagg tttagacgac aaaggcgatg cattgggtca gcagtacaga accgtcgacg
     4441 aagttgtaag tggtggatca gatatcatca ttgttggcag aggacttttc gccaagggta
     4501 gagatcctaa ggttgaaggt gaaagataca gaaatgctgg atgggaagcg taccaaaaga
     4561 gaatcagcgc tccccattaa ttatacagga aacttaatag aacaaatcac atatttaatc
     4621 taatagccac ctgcattggc acggtgcaac actacttcaa cttcatctta caaaaagatc
     4681 acgtgatctg ttgtattggg atctctagac ctaataactt cgtatagcat acattatacg
     4741 aagttatatt aagggttgct cgagtaaacc aattttattt gaacttgccc cgctaatcgt
     4801 acagacgtac cagatccagg tcataacggt gccattagaa ggccgtagaa tagacccact
     4861 aaagctatca caggcaacct tgcatccaag tctaattaaa cggagcagaa tctgttccag
     4921 taatgtataa agaattaact accggaaaag cgtcttccgg ggcggctcaa ccggattaca
     4981 ttcttttttc ttttttgcgg ccagcattcg agtcacgtgc tcaaatactg aggaaggggt
     5041 tcttccggta ggcaccggcg ctcactcttc cgtgcggcac actccccgca gttcaggggc
     5101 catgtgcctg tgagagtatt tccgcaggcc atagttccgg ggtggaccac tccgagctaa
     5161 tgctaaagta agcccttccc aacttgaaat aggaacaaaa gtcaattatc tacctgaatc
     5221 taagattggg ccctagagtg agacgttaat tggtagagtg agacgtttaa gcgggggcgg
     5281 tagagtgaga cgttgggctt agagtgagac gttgccggca atcctcgagc agctcctcct
     5341 gtaaagctaa acgtgtaaag attttagagt gagacgttgt tagcagtcgt tgttcaaggg
     5401 agggatgagt tgaggaataa aatgcggaaa aagtgtgggt tagacagaca tatataaact
     5461 ctagccattc ggctgaaata tatctgagtt catttgtgat aacttttttg attacttgtt
     5521 catatatcca caagaacaac gactattaaa caatagaaaa tagtata
//
