LOCUS       sequence-437583-        8081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Cyterm-yemiRFP670 in budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   Botman D, de Groot DH, Wehrens M, van Rossum K, Yska S, van der 
            Kieft B, Albert V, Minghetti N, Goedhart J, Teusink B
  TITLE     Side-by-side systematic characterization of novel FPs in budding 
            yeast
  JOURNAL   bioRxiv 2025.06.27.661975; doi: 
            https://doi.org/10.1101/2025.06.27.661975
REFERENCE   2  (bases 1 to 8081)
  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: "bioRxiv 
            2025.06.27.661975; doi: https://doi.org/10.1101/2025.06.27.661975"
COMMENT     Sequence Label: pDRF1 Cyterm-yemiRFP670
FEATURES             Location/Qualifiers
     source          1..8081
                     /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"
     regulatory      1024..1033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1971..1995)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2057..2076)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2058..2076)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2224..2411
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2279..2303)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2423..2440)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2423..2439)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2432..2454)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2875..4221
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4332..5135)
                     /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(5136..5351)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5376..5394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5462..5566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5567..5635,5636..6427)
                     /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(5785..5804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6598..7186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7087..7106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7340..7357
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7474..7495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7510..7515,7516..7533,7534..7540)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7548..7564
                     /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     7553..7575
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7572..7588
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7572..7588
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7618..8073)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7755..7776)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7967..7986
                     /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 gctaccggac
      841 tcagatctgg taccccttgc agcttccagt tgtcaggaag agaaggctcc catggatcct
      901 gtggtggtgc tggggctctg tctctcctgt ttgcttctcc tttcactctg gaaacagagc
      961 tatgggggag ggaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     1021 gtcgccacca tggctgaagg ttctgttgct agacaaccag atttgttgac ttgtgaacat
     1081 gaagaaattc atttggctgg ttctattcaa ccacatggtg ctttgttggt cgtttctgaa
     1141 catgaccata gagttattca agcctctgct aatgccgctg aattcttaaa cttgggttct
     1201 gttttaggtg ttccattagc tgaaattgat ggtgatttgt taattaagat tttaccacat
     1261 ttagatccaa ccgctgaagg tatgccagtt gctgttagat gtagaatcgg taatccttcc
     1321 actgaatact gtggtttgat gcatagacca ccagaaggtg gtttgattat tgaattggaa
     1381 agagctggtc catctattga tttatccggt actttggctc cagctttgga aagaattaga
     1441 actgctggtt ctttaagagc tttgtgtgat gacactgttt tgttattcca acaatgtact
     1501 ggttatgata gagttatggt ttacagattc gatgaacaag gtcatggttt agttttctct
     1561 gaatgtcatg tccctggttt ggaatcttac ttcggtaata gatacccatc ctctactgtt
     1621 ccacaaatgg ctagacaatt atacgttaga caaagagtta gagttttggt cgatgttact
     1681 taccaaccag ttccattgga accaagatta tccccattga ctggtagaga tttagatatg
     1741 tctggttgtt tcttaagatc tatgtctcct tgtcatttgc aattcttgaa ggatatgggt
     1801 gttagagcta ctttggccgt ttctttagtt gttggtggta agttgtgggg tttggttgtt
     1861 tgtcaccatt acttgccaag attcattaga ttcgaattaa gagctatctg taagagattg
     1921 gctgaaagaa ttgctactag aattaccgct ttagaatctt aagcggccgc acccagcttt
     1981 cttgtacaaa gtggtgtagt taattcatga tggccgctgc aggtcgacct cgaggggggg
     2041 cccggtaccc aattcgccct atagtgagtc gtattacgcg cggatccagc tttggacttc
     2101 ttcgccagag gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac cttgccagaa
     2161 atttacgaaa agatggaaaa gggtcaaatc gttggtagat acgttgttga cacttctaaa
     2221 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     2281 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     2341 tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac
     2401 acctctaccg gcatgccaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2461 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2521 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2581 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataatcggat
     2641 cgtacttgtt acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca
     2701 gatagtatat ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta
     2761 tgtatttcgg ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca
     2821 tccccggttc attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa
     2881 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     2941 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     3001 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     3061 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     3121 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     3181 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     3241 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     3301 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     3361 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     3421 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     3481 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     3541 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     3601 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     3661 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     3721 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     3781 gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa
     3841 gttcctatac tttctagcta gagaatagga acttcggaat aggaacttca aagcgtttcc
     3901 gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt
     3961 cgcacctata tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg
     4021 tgtttatgct taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag
     4081 tacctcctgt gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc
     4141 ctttagctgt tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat
     4201 catttccttt gatattggat cgatccgatg ataagctgtc aaacatgaga attgggtaat
     4261 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     4321 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     4381 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     4441 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     4501 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     4561 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     4621 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     4681 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     4741 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     4801 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     4861 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     4921 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     4981 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     5041 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     5101 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcatgtt
     5161 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     5221 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     5281 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     5341 gatgaattga aaagctaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     5401 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5461 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5521 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5581 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5641 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5701 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5761 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5821 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5881 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5941 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6001 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6061 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6121 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6181 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6241 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6301 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6361 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6421 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6481 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6541 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6601 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6661 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6721 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6781 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6841 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6901 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6961 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7021 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7081 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7141 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7201 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7261 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7321 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7381 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     7441 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     7501 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     7561 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttaccg catcaggaaa
     7621 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     7681 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     7741 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     7801 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7861 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7921 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7981 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     8041 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg c
//
