LOCUS       sequence-181610-        8554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HO-TEF1p-ATeam1.03-KanMX4-HO. Codon optimized for yeast version of 
            the ATeam1.03 ATP FRET sensor (originally from Imamura et al. PNAS, 
            2009 ), expressed via the TEF1 promoter. HO locus integration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8554)
  AUTHORS   Papagiannakis A, Niebel B, Wit EC, Heinemann M
  TITLE     Autonomous Metabolic Oscillations Robustly Gate the Early and Late 
            Cell Cycle.
  JOURNAL   Mol Cell. 2017 Jan 19;65(2):285-295. doi: 
            10.1016/j.molcel.2016.11.018. Epub 2016 Dec 15.
  PUBMED    27989441
REFERENCE   2  (bases 1 to 8554)
  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"; doi: 
            "10.1016/j.molcel.2016.11.018"; journalName: "Mol Cell"; date: 
            "2017-01-19- 19"; volume: "65"; issue: "2"; pages: "285-295"
COMMENT     Sequence Label: pTEF:ATP
FEATURES             Location/Qualifiers
     source          1..8554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    97..118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    173..189
                     /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)."
     primer_bind     178..200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     197..213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     197..213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(1205..1394)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(3280..3678)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     gene            3761..5117
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        3761..4104
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             4105..4914
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4172..4191)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4920..5117
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5725..5744)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5726..5744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5751..5768)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5751..5767)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5760..5782)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6067..6086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6199..6220
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6374..6393)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6493..6515
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6553..6571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6639..6743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6744..6812,6813..7604)
                     /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     complement(6962..6981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7775..8363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8264..8283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8517..8534
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
       61 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      121 ggcaccccag gctttacact ttatgcttcc gcggctcgta tgttgtgtgg aattgtgagc
      181 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccactag tccgaggcct
      241 cgagatccga tatcgccgtg gcggccgcca gctgaagctt aattatcctg ggcacgagtg
      301 aaacaaagct aaaaccttta tttagcatgg ccattgaatg taacaattat atatatcgca
      361 agcacaaaaa atcaaggaga gagaactacc actttgttca tgtgtacaat gttcattatc
      421 tccataagca aaaaaaaaaa aatagaaaac atatgctata aggttgatat tctcacgagt
      481 aagcggcact tgctacttat tgacattgca gatttttggc tacagaaata gtatattaga
      541 gattataatt gctaatcaaa tcaaaatata aaattagtaa accaaaccat ttataccctt
      601 ccttagtagt tatggattgt tttttaatga tatttctgca aaccaaagaa agattgttat
      661 ccagatagaa tttagttttg atattcattt ttttgttgaa gattgaacgc catatctggg
      721 cctcataatt caaaagacgg tgccattatc ggtagcgttt cgcattgtac tggatttcag
      781 aaatttcaca gttgatgaat cgaaaagaat ggtctcattg caacacgtaa ggttaagatg
      841 tcccttttta ccattatagg caataaatga atcataaaac gaccgtatac tggtgaaata
      901 gtagggagaa cgagtacctg tagtaaaaag tataaatcat agttaatcgg gcaatgtccc
      961 tcgatcaagg agtattgtgt catgttcgag acaaacgcca acatttttgt ttcttttgga
     1021 caaatgttgt ttgcatttat gatccgttat attttgatct aatgtagagt tgcacgtagt
     1081 tcttactggc aaagaaatcg atgcatacca aaaaagaata aaggtgatat ttgatcttta
     1141 ccgtttagtt ccaacgtaaa attgtgcctt tggacttaaa atggcgtcgt acgctgagct
     1201 ggatcttcga gcgtcccaaa accttctcaa gcaaggtttt cagtataatg ttacatgcgt
     1261 acacgcgtct gtacagaaaa aaaagaaaaa tttgaaatat aaataacgtt cttaatacta
     1321 acataactat aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt
     1381 cggttagagc ggatcttagc tagccgcggt accaagctta ctcgagtcat tcaatgttat
     1441 gtctaatttt gaagttggcc ttgataccat ttttttgttt gtcagcagtg atgtagacgt
     1501 tgtgagagtt ataattgtat tctaacttat gacctaaaat gttaccatct tctttaaaat
     1561 caataccttt caattcaatt ctattgacca aagtgtcacc ttcaaatttt acttcagctc
     1621 tggtcttgta attaccatcg tccttaaaga aaatagttct ttcttggaca taaccttcag
     1681 gcattgcaga cttaaaaaag tcatgttgct tcatgtggtc aggatatcta gcgaagcatt
     1741 gtaaaccgta acccaatgtt gtaactaatg taggccaagg aacaggtaac ttaccggtag
     1801 tgcatatcaa tttcaaagtc aatttaccgt atgttgcgtc accttcacct tcaccactaa
     1861 ctgaaaactt atgaccattg acgtcaccgt ctaattctac taaaataggg acgacacctg
     1921 tgaataattc ttcaccttta ctaaccatac caccagaacc acccttgtat aattcatcca
     1981 tacccaaagt aataccggca gcggtgacaa attctaataa aaccatatga tctctttttt
     2041 cgttaggatc tttggacaac ttagattggt aggacaaata gtgattatca ggtaataata
     2101 caggaccgtc accgataggt gtgttttgtt gatagtggtc tgctaattga acaccaccgt
     2161 ccatgaattc gttagcctta ccagcaacat ccaatctgtt caaagctctt tgtaaagcca
     2221 attctgctct tctgatatcg gtatcatcag attgagagtt caatctttct tgggctcttt
     2281 gtctagcagc ttcagctctt tctttgtcaa taccttcagc tgtttcagca gcttgagcgt
     2341 taatagtaac atgatctggt ctaacttcaa cagtaccacc tgaaactgca accatttctg
     2401 tttgaccgtc ctttttcaat ctaacggcac caattttcaa tggggctttt gtaggaatat
     2461 gacctggcaa aatacccaaa tcaccagatt cagctctgac agaaaccatt tcaatatcgg
     2521 catcgtaaac aggaccatct ggagtagtaa tgttaacctt aacggtcttc atatcgatag
     2581 cagcagtaac aaattccaac aagaccatgt gatctctttt ttcgtttggg tccttagaca
     2641 atttggattg ggtagacaag taatggttat ctggcaacaa aactggacca tcaccaattg
     2701 gagtgttttg ttggtaatga tcagccaatt gtacaccacc atcttcgatg ttgtgtctga
     2761 tcttgaaatg agccttaata ccgttctttt gcttatcggc ggtaatgtaa acgttatggg
     2821 agatgtagtt gtattccaac ttgtgaccca agatgttacc atcttctttg aagtcgatac
     2881 ccttcaattc gattctgtta accaaggtat caccttcgaa cttaacttcg gctctagttt
     2941 tgtagttacc gtcatccttg aagaagatgg ttctttcttg aacgtaacct tctggcatag
     3001 cggatttgaa gaaatcgtgt tgcttcatat gatctgggta tctagcaaaa cattgaacac
     3061 cccaagtcaa agtagtaacc aaagttggcc atggaactgg caatttacct gtggtacaga
     3121 tgaacttcaa ggtcaattta ccataagtag catcaccttc accttcacca gaaacagaaa
     3181 atctgtgacc gttaacatca ccatccaatt caaccaagat tggaacaaca ccagtgaata
     3241 attcttcacc cttagaaacc attttaataa cctaggaaac ttagattaga ttgctatgct
     3301 ttctttctaa tgagcaagaa gtaaaaaaag ttgtaataga acaagaaaaa tgaaactgaa
     3361 acttgagaaa ttgaagaccg tttattaact taaatatcaa tgggaggtca tcgaaagaga
     3421 aaaaaatcaa aaaaaaaatt ttcaagaaaa agaaacgtga taaaaatttt tattgccttt
     3481 ttcgacgaag aaaaagaaac gaggcggtct cttttttctt ttccaaacct ttagtacggg
     3541 taattaacga caccctagag gaagaaagag gggaaattta gtatgctgtg cttgggtgtt
     3601 ttgaagtggt acggcgatgc gcggagtccg agaaaatctg gaagagtaaa aaaggagtag
     3661 aaacattttg aagctatggt gtggaattcc agctgcgggt taattaaggc gcgccagatc
     3721 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
     3781 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
     3841 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
     3901 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
     3961 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     4021 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     4081 catcacatcc gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga
     4141 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     4201 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     4261 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     4321 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     4381 tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct
     4441 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     4501 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     4561 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     4621 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc
     4681 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     4741 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     4801 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     4861 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta
     4921 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     4981 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     5041 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     5101 tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga
     5161 gctcgaatta attcctgggg gaacaacttc acagaatgtt ttgtcatatt gtcgaagtgg
     5221 tcacaaaaca agagaagttc cgccaattat aaaaagggaa cccgtatatt tcagcttcac
     5281 ggatgatttc cagggtgaga gtactgtata tgggcttacg atagaaggcc ataaaaattt
     5341 cttgcttggc aacaaaatag aagtgaaatc atgtcgaggc tgctgtgtgg gagaacagca
     5401 taaaatatca caaaaaaaga atctaaaaca ctgtgttgct tgtcccagaa agggaatcaa
     5461 gtatttttat aaagattgga gtggtaaaaa tcgagtatgt gctagatgct atggaagata
     5521 caaattcagc ggtcatcact gtataaattg caagtatgta ccagaagcac gtgaagtgaa
     5581 aaaggcaaaa gacaaaggcg aaaaattggg cattacgccc gaaggtttgc cagttaaagg
     5641 accagagtgt ataaaatgtg gcggaatctt acagtggcct atgcggccgc tctagctaga
     5701 actagtggat cgatccccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     5761 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5821 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5881 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     5941 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     6001 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     6061 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6121 aagctctaaa tcggggtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6181 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     6241 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     6301 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     6361 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     6421 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6481 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     6541 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     6601 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6661 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6721 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6781 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6841 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6901 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6961 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7021 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7081 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7141 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7201 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7261 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7321 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7381 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7441 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7501 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7561 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7621 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7681 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7741 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7801 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7861 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7921 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7981 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8041 ccgggttgga ctcaagacga tagttaccgg ataaggcaca gcggtcgggc tgaacggggg
     8101 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8161 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8221 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8281 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8341 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8401 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8461 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8521 agtcagtgag cgaggaagcg gaagagcgcc caat
//
