LOCUS       sequence-370761-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the TEF1 promoter appended with 3 tetracycline riboswitch 
            using a green fluorescence protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  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 5930)
  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: pILGFP3A2 (2)
FEATURES             Location/Qualifiers
     source          1..5930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             15..728
                     /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      738..1008
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1273..1289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1273..1289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1286..1308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1297..1313
                     /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(1321..1327,1328..1345,1346..1351))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1366..1387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1504..1521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1675..2263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1755..1774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2434..3225,3226..3294))
                     /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     3057..3076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3295..3399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3467..3485
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3523..3545)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3645..3664
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3858..3880
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3872..3889
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3873..3889
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5003..5036)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5256..5669
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccacctaggc caaaatgtct aaaggtgaag aattattcac tggtgttgtc ccaattttgg
       61 ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg
      121 atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa ttgccagttc
      181 catggccaac cttagtcact actttcggtt atggtgttca atgttttgcg agatacccag
      241 atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat gttcaagaaa
      301 gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc aagtttgaag
      361 gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa gatggtaaca
      421 ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc atggctgaca
      481 aacaaaagaa tggtatcaaa gttaacttca aaattagaca caacattgaa gatggttctg
      541 ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac
      601 cagacaacca ttacttatcc actcaatctg ccttatccaa agatccaaac gaaaagagag
      661 accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt atggatgaat
      721 tgtacaaaag atcttaaatt gaattgaatt gaaatcgata gatcaatttt tttcttttct
      781 ctttccccat cctttacgct aaaataatag tttattttat tttttgaata ttttttattt
      841 atatacgtat atatagacta ttatttatct tttaatgatt attaagattt ttattaaaaa
      901 aaaattcgct cctcttttaa tgcctttatg cagttttttt ttcccattcg atatttctat
      961 gttcgggttc agcgtatttt aagtttaata actcgaaaat tctgcgttcg ttaaagcttt
     1021 cgagaaggat attatttact agtaggacta tttgcaaagg gaagggatgc taaggtagag
     1081 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     1141 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     1201 taattatatc agttattacc cgggaatctc ggtcgtaatg atttaaatgc atgcaagctt
     1261 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1321 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1381 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1441 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2101 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2341 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2401 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2461 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2521 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2581 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2641 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2701 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2761 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2821 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2881 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2941 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3001 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3061 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3121 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3181 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3241 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3301 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3361 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3421 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3481 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3541 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3601 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3661 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3721 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     3781 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     3841 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcattta
     3901 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     3961 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     4021 acccaactgc acagaacaaa aacctgcagg aaacgaagat aaaacatgtc cacaaaatca
     4081 tataccagta gagctgagac tcatgcaagt ccggttgcat cgaaactttt acgtttaatg
     4141 gatgaaaaga agaccaattt gtgtgcttct cttgacgttc gttcgactga tgagctattg
     4201 aaacttgttg aaacgttggg tccatacatt tgccttttga aaacacacgt tgatatcttg
     4261 gatgatttca gttatgaggg tactgtcgtt ccattgaaag cattggcaga gaaatacaag
     4321 ttcttgatat ttgaggacag aaaattcgcc gatatcggta acacagtcaa attacaatat
     4381 acatcgggcg tttaccgtat cgcagaatgg tctgatatca ccaacgccca cggggttact
     4441 ggtgctggta ttgttgctgg cttgaaacaa ggtgcgcaag aggtcaccaa agaaccaagg
     4501 ggattattga tgcttgctga attgtcttcc aagggttctc tagcacacgg tgaatatact
     4561 aagggtaccg ttgatattgc aaagagtgat aaagatttcg ttattgggtt cattgctcag
     4621 aacgatatgg gaggaagaga agaagggttt gattggctaa tcatgacccc aggtgtaggt
     4681 ttagacgaca aaggcgatgc attgggtcag cagtacagaa ccgtcgacga agttgtaagt
     4741 ggtggatcag atatcatcat tgttggcaga ggacttttcg ccaagggtag agatcctaag
     4801 gttgaaggtg aaagatacag aaatgctgga tgggaagcgt accaaaagag aatcagcgct
     4861 ccccattaat tatacaggaa acttaataga acaaatcaca tatttaatct aatagccacc
     4921 tgcattggca cggtgcaaca ctacttcaac ttcatcttac aaaaagatca cgtgatctgt
     4981 tgtattggga tctctagacc taataacttc gtatagcata cattatacga agttatatta
     5041 agggttgctc gagaaagagc tccagaaagc gaccacccaa ctttggctga taatagcgta
     5101 taaacaatgc atactttgta cgttcaaaat acaatgcagt agatatattt atgcatatta
     5161 catataatac atatcacata ggaagcaaca ggcgcgttgg acttttaatt ttcgaggacc
     5221 gcgaatcctt acatcacacc caatccccca caagtgatcc cccacacacc atagcttcaa
     5281 aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc
     5341 acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg
     5401 ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt
     5461 cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt
     5521 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
     5581 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
     5641 agaaagaaag catagcaatc taatctaagt tttaattaca aacggcctaa aacataccag
     5701 atcgccaccc gcgctttaat ctggagaggt gaagaatacg accacctagg ccaactcgca
     5761 aatcaacaac tctcaattca aggcctaaaa cataccagat cgccacccgc gctttaatct
     5821 ggagaggtga agaatacgac cacctaggcc aactcgcaaa tcaacaactc gaggcctaaa
     5881 acataccaga tcgccacccg cgctttaatc tggagaggtg aagaatacga
//
