LOCUS       sequence-369622-        5780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the TEF1 promoter inserted with 4 Z268 elements using a 
            green fluorescence protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5780)
  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 5780)
  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: pILGFP4D42
FEATURES             Location/Qualifiers
     source          1..5780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             301..1014
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1024..1294
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1559..1575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1559..1575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1572..1594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1583..1599
                     /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(1607..1613,1614..1631,1632..1637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1652..1673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1790..1807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1961..2549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2041..2060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2720..3511,3512..3580))
                     /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     3343..3362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3581..3685)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3753..3771
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3809..3831)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3931..3950
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4144..4166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4158..4175
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4159..4175
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5289..5322)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ctacctgaat ctaagattgg gcccgcgtgg gcgaattggt gcgtgggcgt aagcgggggc
       61 gggcgtgggc gtagcgtggg cggccggcaa tcctcgagca gattcttttt cttgaaaatt
      121 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
      181 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
      241 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa aggatccaaa
      301 atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga attagatggt
      361 gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc tacttacggt
      421 aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg gccaacctta
      481 gtcactactt tcggttatgg tgttcaatgt tttgcgagat acccagatca tatgaaacaa
      541 catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac tatttttttc
      601 aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga taccttagtt
      661 aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt aggtcacaaa
      721 ttggaataca actataactc tcacaatgtt tacatcatgg ctgacaaaca aaagaatggt
      781 atcaaagtta acttcaaaat tagacacaac attgaagatg gttctgttca attagctgac
      841 cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga caaccattac
      901 ttatccactc aatctgcctt atccaaagat ccaaacgaaa agagagacca catggtcttg
      961 ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta caaaagatct
     1021 taaattgaat tgaattgaaa tcgatagatc aatttttttc ttttctcttt ccccatcctt
     1081 tacgctaaaa taatagttta ttttattttt tgaatatttt ttatttatat acgtatatat
     1141 agactattat ttatctttta atgattatta agatttttat taaaaaaaaa ttcgctcctc
     1201 ttttaatgcc tttatgcagt ttttttttcc cattcgatat ttctatgttc gggttcagcg
     1261 tattttaagt ttaataactc gaaaattctg cgttcgttaa agctttcgag aaggatatta
     1321 tttactagta ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     1381 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     1441 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     1501 attacccggg aatctcggtc gtaatgattt aaatgcatgc aagcttggcg taatcatggt
     1561 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1621 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1681 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1741 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2401 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3661 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     3721 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3781 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3841 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3901 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3961 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     4021 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4081 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4141 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt catttaaatt tttttgattc
     4201 ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat
     4261 acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag
     4321 aacaaaaacc tgcaggaaac gaagataaaa catgtccaca aaatcatata ccagtagagc
     4381 tgagactcat gcaagtccgg ttgcatcgaa acttttacgt ttaatggatg aaaagaagac
     4441 caatttgtgt gcttctcttg acgttcgttc gactgatgag ctattgaaac ttgttgaaac
     4501 gttgggtcca tacatttgcc ttttgaaaac acacgttgat atcttggatg atttcagtta
     4561 tgagggtact gtcgttccat tgaaagcatt ggcagagaaa tacaagttct tgatatttga
     4621 ggacagaaaa ttcgccgata tcggtaacac agtcaaatta caatatacat cgggcgttta
     4681 ccgtatcgca gaatggtctg atatcaccaa cgcccacggg gttactggtg ctggtattgt
     4741 tgctggcttg aaacaaggtg cgcaagaggt caccaaagaa ccaaggggat tattgatgct
     4801 tgctgaattg tcttccaagg gttctctagc acacggtgaa tatactaagg gtaccgttga
     4861 tattgcaaag agtgataaag atttcgttat tgggttcatt gctcagaacg atatgggagg
     4921 aagagaagaa gggtttgatt ggctaatcat gaccccaggt gtaggtttag acgacaaagg
     4981 cgatgcattg ggtcagcagt acagaaccgt cgacgaagtt gtaagtggtg gatcagatat
     5041 catcattgtt ggcagaggac ttttcgccaa gggtagagat cctaaggttg aaggtgaaag
     5101 atacagaaat gctggatggg aagcgtacca aaagagaatc agcgctcccc attaattata
     5161 caggaaactt aatagaacaa atcacatatt taatctaata gccacctgca ttggcacggt
     5221 gcaacactac ttcaacttca tcttacaaaa agatcacgtg atctgttgta ttgggatctc
     5281 tagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg ttgctcgaga
     5341 aagagctcca gaaagcgacc acccaacttt ggctgataat agcgtataaa caatgcatac
     5401 tttgtacgtt caaaatacaa tgcagtagat atatttatgc atattacata taatacatat
     5461 cacataggaa gcaacaggcg cgttggactt ttaattttcg aggaccgcga atccttacat
     5521 cacacccaat cccccacaag tgatccccca cacaccatag cttcaaaatg tttctactcc
     5581 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
     5641 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
     5701 taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca
     5761 ataaaaattt ttatcacgtt
//
