LOCUS       sequence-326046-        8421 bp    DNA     circular SYN 15-APR-2025
DEFINITION  FRET fusion between ymTq2 and AausFP1 for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8421)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: ymTq2-AausFP1 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2311..2335)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2397..2416)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2398..2416)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2564..2751
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2619..2643)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2763..2780)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2763..2779)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2772..2794)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3215..4561
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4672..5475)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5476..5691)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5716..5734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5802..5906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5907..5975,5976..6767)
                     /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(6125..6144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6938..7526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7427..7446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7680..7697
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7814..7835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7850..7855,7856..7873,7874..7880)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7888..7904
                     /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     7893..7915
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7912..7928
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7912..7928
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7958..8413)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8095..8116)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8307..8326
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc actagtaagc
      841 ttttaattaa atggttagta aaggtgaaga attgttcact ggtgtagtgc caattttagt
      901 agaattagat ggtgacgtta acggtcataa gttctctgta agcggtgaag gtgaaggtga
      961 cgcaacatac ggtaaattaa ctttgaaatt catttgtact acaggtaaat taccagttcc
     1021 atggccaact ttggttacta ctttgtcttg gggtgttcaa tgtttcgcta gatacccaga
     1081 tcatatgaag caacatgatt tctttaagtc tgctatgcca gaaggttacg ttcaagaaag
     1141 aactatcttc ttcaaggatg atggtaacta taagactaga gctgaagtta agttcgaagg
     1201 agatacttta gttaatagaa ttgaattgaa gggtattgat tttaaggaag atggtaatat
     1261 tttgggtcat aagttggaat ataattactt ctctgataac gtttacatta ctgctgataa
     1321 gcaaaagaac ggtattaagg cgaacttcaa gattagacat aacattgaag atggtggtgt
     1381 tcaattagct gatcattacc aacaaaacac tccaattggt gacggtccag ttttgttgcc
     1441 agataaccat tacttgtcta ctcaatctaa gttgtctaag gatccaaacg aaaagagaga
     1501 tcatatggtt ttgttggaat tcgttactgc tgctgggatc actttaggta tggatgaatt
     1561 gtataaatcc ggactcagat cttcggatcc accggtcgcc accatgagtt acggagcact
     1621 tttgttcaga gaaaaaatcc cctatgttgt tgagatggaa ggtgatgttg agggaatgaa
     1681 gttttcagta agaggtaaag gacatggtga tgctaacact ggtaaaattg aagccagttt
     1741 catctgcaca actggtgaac taccagtacc ttggtcaagc attctaacaa ctgtcacata
     1801 cggtgcacag tgttttgcaa aatatccaaa tgatatcaaa gactacccaa aaagtgctat
     1861 gccagaaggt tacgttcaag agcgaacaat cacttttgaa aacgacggcg tttataagac
     1921 acgcgctgaa gtcacgtacg agaaaggatc agtttataac cgagttacgt tgaacggttc
     1981 aggatttaaa aaaggcggta acatcttggg caaaaaattg gaatttaact acaaccctca
     2041 ttgcatctat gttcttccag atgtacaaaa taatggaatc aaatgctaca tcaacattgt
     2101 tcatgacgta attggaggag gacaaataat tgctgcgcac caacaattaa atactcctct
     2161 tggcggtgga ccagtcgaca ttccacacta ccatcatata caagcgcaca caatactttc
     2221 taaagatccc aaagaaacaa gagatcatat gaacgtagta gaggtgttca gagctatcga
     2281 ttgcaaaacc gcatatgcat aagcggccgc acccagcttt cttgtacaaa gtggtgtagt
     2341 taattcatga tggccgctgc aggtcgacct cgaggggggg cccggtaccc aattcgccct
     2401 atagtgagtc gtattacgcg cggatccagc tttggacttc ttcgccagag gtttggtcaa
     2461 gtctccaatc aaggttgtcg gcttgtctac cttgccagaa atttacgaaa agatggaaaa
     2521 gggtcaaatc gttggtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt
     2581 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2641 ctcttaggtt ttaaaacgan aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2701 gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccaat
     2761 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2821 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2881 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2941 cttacgcatc tgtgcggtat ttcacaccgc ataatcggat cgtacttgtt acccatcatt
     3001 gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat ttgaacctgt
     3061 ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg ttcctggaga
     3121 aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc attttctgcg
     3181 tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc ttcattttgt
     3241 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     3301 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     3361 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     3421 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     3481 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     3541 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     3601 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     3661 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     3721 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3781 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3841 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3901 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3961 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     4021 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     4081 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     4141 gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     4201 gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa
     4261 atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt
     4321 gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta
     4381 cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
     4441 cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
     4501 gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
     4561 cgatccgatg ataagctgtc aaacatgaga attgggtaat aactgatata attaaattga
     4621 agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg
     4681 ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac
     4741 cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt
     4801 agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa
     4861 acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct
     4921 ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt
     4981 agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc
     5041 tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc
     5101 caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga
     5161 gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa
     5221 attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt
     5281 caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc
     5341 cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg
     5401 catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata
     5461 tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt
     5521 taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct
     5581 aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt
     5641 tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctaatt
     5701 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5761 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5821 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5881 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5941 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6001 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6061 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6121 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6181 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6241 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6301 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6361 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6421 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6481 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6541 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6601 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6661 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6721 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6781 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6841 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6901 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6961 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7021 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7081 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7141 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7201 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7261 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7321 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7381 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7441 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7501 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7561 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7621 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7681 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7741 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7801 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     7861 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     7921 gctatgacca tgattacgcc aagcttaccg catcaggaaa ttgtaagcgt taatattttg
     7981 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     8041 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     8101 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     8161 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     8221 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     8281 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     8341 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     8401 ctacagggcg cgtccattcg c
//
