LOCUS       sequence-370799-        5545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the SkGAL2 promoter inserted with 4 Z268 elements using a 
            green fluorescent protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5545)
  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 5545)
  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: pILGFP10CF71
FEATURES             Location/Qualifiers
     source          1..5545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             836..1546
                     /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(1791..1807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1791..1807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1804..1826)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1815..1831
                     /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(1839..1845,1846..1863,1864..1869))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1884..1905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2022..2039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2193..2781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2273..2292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2952..3743,3744..3812))
                     /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     3575..3594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3813..3917)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3985..4003
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4041..4063)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4163..4182
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4376..4398
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4390..4407
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4391..4407
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(5521..5545,1..9))
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cgaagttata ttaagggttg ctcgagtaaa ccaattttat ttgaacttgc cccgctaatc
       61 gtacagacgt accagatcca ggtcataacg gtgccattag aaggccgtag aatagaccca
      121 ctaaagctat cacaggcaac cttgcatcca agtctaatta aacggagcag aatctgttcc
      181 agtaatgtat aaagaattaa ctaccggaaa agcgtcttcc ggggcggctc aaccggatta
      241 cattcttttt tcttttttgc ggccagcatt cgagtcacgt gctcaaatac tgaggaaggg
      301 gttcttccgg taggcaccgg cgctcactct tccgtgcggc acactccccg cagttcaggg
      361 gccatgtgcc tgtgagagta tttccgcagg ccatagttcc ggggtggacc actccgagct
      421 aatgctaaag taagcccttc ccaacttgaa ataggaacaa aagtcaatta tctacctgaa
      481 tctaagattg ggcccgcgtg ggcgaattgg tgcgtgggcg taagcggggg cgggcgtggg
      541 cgtagcgtgg gcggccggca atcctcgagc agctcctcct gtaaagctaa acgtgtaaag
      601 attttagagt gagacgttgt tagcagtcgt tgttcaaggg agggatgagt tgaggaataa
      661 aatgcggaaa aagtgtgggt tagacagaca tatataaact ctagccattc ggctgaaata
      721 tatctgagtt catttgtgat aacttttttg attacttgtt catatatcca caagaacaac
      781 gactattaaa caatagaaaa tagtataata aaaaaagttt tttacaatgg gatcctctaa
      841 aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa
      901 tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac
      961 cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac
     1021 tttcggttat ggtgttcaat gttttgcgag atacccagat catatgaaac aacatgactt
     1081 tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga
     1141 cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat
     1201 cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata
     1261 caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt
     1321 taacttcaaa attagacaca acattgaaga tggttctgtt caattagctg accattatca
     1381 acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac
     1441 tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt
     1501 tgttactgct gctggtatta cccatggtat ggatgaattg tacaaaagat cttaaactag
     1561 taggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     1621 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     1681 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     1741 ggaatctcgg tcgtaatgat ttaaatgcat gcaagcttgg cgtaatcatg gtcatagctg
     1801 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1861 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1921 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1981 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2041 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2101 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2161 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2221 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2281 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2341 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2401 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2461 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2521 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2581 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2641 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2701 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2821 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2881 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2941 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3001 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3061 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3121 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3181 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3241 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3301 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3361 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3421 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3481 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3541 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3601 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3661 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3721 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3781 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3841 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3901 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3961 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4021 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4081 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4141 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     4261 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4321 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     4381 tcacgacgtt gtaaaacgac ggccagtgaa ttcatttaaa tttttttgat tcggtaatct
     4441 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     4501 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     4561 cctgcaggaa acgaagataa aacatgtcca caaaatcata taccagtaga gctgagactc
     4621 atgcaagtcc ggttgcatcg aaacttttac gtttaatgga tgaaaagaag accaatttgt
     4681 gtgcttctct tgacgttcgt tcgactgatg agctattgaa acttgttgaa acgttgggtc
     4741 catacatttg ccttttgaaa acacacgttg atatcttgga tgatttcagt tatgagggta
     4801 ctgtcgttcc attgaaagca ttggcagaga aatacaagtt cttgatattt gaggacagaa
     4861 aattcgccga tatcggtaac acagtcaaat tacaatatac atcgggcgtt taccgtatcg
     4921 cagaatggtc tgatatcacc aacgcccacg gggttactgg tgctggtatt gttgctggct
     4981 tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg attattgatg cttgctgaat
     5041 tgtcttccaa gggttctcta gcacacggtg aatatactaa gggtaccgtt gatattgcaa
     5101 agagtgataa agatttcgtt attgggttca ttgctcagaa cgatatggga ggaagagaag
     5161 aagggtttga ttggctaatc atgaccccag gtgtaggttt agacgacaaa ggcgatgcat
     5221 tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg tggatcagat atcatcattg
     5281 ttggcagagg acttttcgcc aagggtagag atcctaaggt tgaaggtgaa agatacagaa
     5341 atgctggatg ggaagcgtac caaaagagaa tcagcgctcc ccattaatta tacaggaaac
     5401 ttaatagaac aaatcacata tttaatctaa tagccacctg cattggcacg gtgcaacact
     5461 acttcaactt catcttacaa aaagatcacg tgatctgttg tattgggatc tctagaccta
     5521 ataacttcgt atagcataca ttata
//
