LOCUS       sequence-159500-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of TEF1 promoter with 
            yEGFP-CLN2PEST (destabilized GFP) used as the reporter gene. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  AUTHORS   Peng B, Williams TC, Henry M, Nielsen LK, Vickers CE
  TITLE     Controlling heterologous gene expression in yeast cell factories on 
            different carbon substrates and across the diauxic shift: a 
            comparison of yeast promoter activities.
  JOURNAL   Microb Cell Fact. 2015 Jun 26;14:91. doi: 10.1186/s12934-015-0278-5.
  PUBMED    26112740
REFERENCE   2  (bases 1 to 5898)
  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: "Microb Cell
            Fact."; date: "2015-06-26"; volume: "14:91. doi"; pages: " 
            10.1186/s12934-015-0278-5"
COMMENT     Sequence Label: pITGFP89
FEATURES             Location/Qualifiers
     source          1..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..36)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(20..36)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(33..55)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    44..60
                     /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(68..74,75..92,93..98))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(251..268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(422..1010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(502..521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1181..1972,1973..2041))
                     /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     1804..1823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2042..2146)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2214..2232
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2270..2292)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2392..2411
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2605..2627
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2619..2636
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2620..2636
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3750..3783)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3993..4406
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4429..5139
                     /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"
ORIGIN
        1 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
       61 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      121 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      301 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      361 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      601 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      841 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
      901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1081 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2161 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2221 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2281 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2341 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2401 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2461 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2521 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2581 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2641 tcatttaaat ttttttgatt cggtaatctc cgagcagaag gaagaacgaa ggaaggagca
     2701 cagacttaga ttggtatata tacgcatatg tggtgttgaa gaaacatgaa attgcccagt
     2761 attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac
     2821 aaaatcatat accagtagag ctgagactca tgcaagtccg gttgcatcga aacttttacg
     2881 tttaatggat gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt cgactgatga
     2941 gctattgaaa cttgttgaaa cgttgggtcc atacatttgc cttttgaaaa cacacgttga
     3001 tatcttggat gatttcagtt atgagggtac tgtcgttcca ttgaaagcat tggcagagaa
     3061 atacaagttc ttgatatttg aggacagaaa attcgccgat atcggtaaca cagtcaaatt
     3121 acaatataca tcgggcgttt accgtatcgc agaatggtct gatatcacca acgcccacgg
     3181 ggttactggt gctggtattg ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga
     3241 accaagggga ttattgatgc ttgctgaatt gtcttccaag ggttctctag cacacggtga
     3301 atatactaag ggtaccgttg atattgcaaa gagtgataaa gatttcgtta ttgggttcat
     3361 tgctcagaac gatatgggag gaagagaaga agggtttgat tggctaatca tgaccccagg
     3421 tgtaggttta gacgacaaag gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt
     3481 tgtaagtggt ggatcagata tcatcattgt tggcagagga cttttcgcca agggtagaga
     3541 tcctaaggtt gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc aaaagagaat
     3601 cagcgctccc cattaattat acaggaaact taatagaaca aatcacatat ttaatctaat
     3661 agccacctgc attggcacgg tgcaacactc acttcaactt catcttacaa aagatcacgt
     3721 gatctgttgt attgggatct ctagacctaa taacttcgta tagcatacat tatacgaagt
     3781 tatattaagg gttgctcgac agaaagcgac cacccaactt tggctgataa tagcgtataa
     3841 acaatgcata ctttgtacgt tcaaaataca atgcagtaga tatatttatg catattacat
     3901 ataatacata tcacatagga agcaacaggc gcgttggact tttaattttc gaggaccgcg
     3961 aatccttaca tcacacccaa tcccccacaa gtgatccccc acacaccata gcttcaaaat
     4021 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     4081 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
     4141 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     4201 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
     4261 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
     4321 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
     4381 aagaaagcat agcaatctaa tctaagtttt aattacaaaa tgggatcctc taaaggtgaa
     4441 gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac
     4501 aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa
     4561 tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttcggt
     4621 tatggtgttc aatgttttgc gagataccca gatcatatga aacaacatga ctttttcaag
     4681 tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac
     4741 tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta
     4801 aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga atacaactat
     4861 aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc
     4921 aaaattagac acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat
     4981 actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct
     5041 gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact
     5101 gctgctggta ttacccatgg tatggatgaa ttgtacaaag catccaactt gaacatttcg
     5161 agaaagctta ccatatcaac cccatcatgc tctttcgaaa attcaaatag cacatccatt
     5221 ccttcgcccg cttcctcatc tcaaagccac actccaatga gaaacatgag ctcactctct
     5281 gataacagcg ttttcagccg gaatatggaa caatcatcac caatcactcc aagtatgtac
     5341 caatttggtc agcagcagtc aaacagtata tgtggtagca ccgttagtgt gaatagtctg
     5401 gtgaatacaa ataacaaaca aaggatctac gaacaaatca cgggtcctaa cagcaataac
     5461 gcaaccaatg attatattga tttgctaaac ctaaatgagt ctaacaagga aaaccaaaat
     5521 cccgcaacgg cgcattacct caatgggggc ccacccaaga caagcttcat taaccatgga
     5581 atgttcccct cgccaactgg gaccataaat agcggtaaat ctagcagtgc ctcatcttta
     5641 atttcttttg gtatgggcaa tacccaagta ataagatctt agactagtag gactatttgc
     5701 aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata
     5761 tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact
     5821 aaactcacaa attagagctt caatttaatt atatcagtta ttacccggga atctcggtcg
     5881 taatgattta aatgcatg
//
