LOCUS       sequence-408442-        7848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-AausFP1 to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7848)
  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 7848)
  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: Cyterm-AausFP1 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      580..1035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(667..686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     877..898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1065..1081)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1065..1081)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1078..1100)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1089..1105
                     /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(1113..1119,1120..1137,1138..1143))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1158..1179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1296..1313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1467..2055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1547..1566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2226..3017,3018..3086))
                     /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     2849..2868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3087..3191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3259..3277
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3302..3517
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3518..4321
                     /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"
     rep_origin      4432..5778
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6199..6221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6213..6230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6214..6230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(6242..6429)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6350..6374
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     6577..6596
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6577..6595
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6658..6682
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7595..7619)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gaaattagat gttagacgat aatgatagga catttgaaaa taatacaata tttggccttt
       61 ttatataatc gaaactaatg gaggggagca taagcggtac ccaccatcga ccacgtttcg
      121 tcaccggtca taattggaat aaacaaaaca aacaaacgaa ctaaagagtg tgagagtaac
      181 aatttccaca gaggtnnana aaaaaaaaaa ggaaaacaga aacaaaaaaa gctcatagtt
      241 ctactatagc ttcttcagac tttctgtgaa cgagggaaaa agacaacaaa aaagcaatgg
      301 ataaacggtc tcgatcgagg cctggaagtg ccgcattagg aaatacagat gcaatgcgcg
      361 cgagtttacg aaattctcgc aggagtttct ccggccatgt gcggctggtt ctacgtctcg
      421 tgtatgggtt tatttctgcg gtaattatcg ccggatctcc gcttcgagac cgggtttgtt
      481 ttcaaattac cggaaaaggt caggcaaaac agagtgatgt atgggtgatc ccagcaatgc
      541 ctgtttatgt ttctccgttt caggaagctt ggcgaatgga cgcgccctgt agcggcgcat
      601 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      661 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      721 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      781 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      841 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      901 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      961 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1021 taacgcttac aatttcctga tgcggtaagc ttggcgtaat catggtcata gctgtttcct
     1081 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1141 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1261 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2101 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2161 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2221 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2281 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2341 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2401 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2461 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2521 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2581 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2641 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2701 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2761 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2821 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2881 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2941 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3001 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3061 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3121 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3181 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3241 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattagct
     3301 tttcaattca tcattttttt tttattcttt tttttgattt cggtttcctt gaaatttttt
     3361 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     3421 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     3481 gcacagaaca aaaacatgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     3541 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     3601 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     3661 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     3721 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     3781 cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     3841 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     3901 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     3961 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     4021 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     4081 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     4141 caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc
     4201 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     4261 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     4321 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     4381 attatatcag ttattaccca attctcatgt ttgacagctt atcatcggat cgatccaata
     4441 tcaaaggaaa tgatagcatt gaaggatgag actaatccaa ttgaggagtg gcagcatata
     4501 gaacagctaa agggtagtgc tgaaggaagc atacgatacc ccgcatggaa tgggataata
     4561 tcacaggagg tactagacta cctttcatcc tacataaata gacgcatata agtacgcatt
     4621 taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag gcaacacgca
     4681 gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa
     4741 gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc tctagctaga
     4801 aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa
     4861 aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac
     4921 aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct
     4981 acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt
     5041 ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa
     5101 tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa
     5161 gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac
     5221 aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa
     5281 atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta
     5341 tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa
     5401 aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg
     5461 ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga
     5521 ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca
     5581 gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc
     5641 acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca
     5701 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg
     5761 aagcacagat gcttcgttaa caaagatatg ctattgaagt gcaagatgga aacgcagaaa
     5821 atgaaccggg gatgcgacgt gcaagattac ctatgcaata gatgcaatag tttctccagg
     5881 aaccgaaata catacattgt cttccgtaaa gcgctagact atatattatt atacaggttc
     5941 aaatatacta tctgtttcag ggaaaactcc caggttcgga tgttcaaaat tcaatgatgg
     6001 gtaacaagta cgatccgatt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6061 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     6181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattggcat
     6241 gccggtagag gtgtggtcaa taagagcgac ctcatgctat acctgagaaa gcaacctgac
     6301 ctacaggaaa gagttactca agaataagaa ttttcgtttt aaaacctaag agtcacttta
     6361 aaatttgtat acacttattt tttttataac ttatttaata ataaaaatca taaatcataa
     6421 gaaattcgct tatttagaag tgtcaacaac gtatctacca acgatttgac ccttttccat
     6481 cttttcgtaa atttctggca aggtagacaa gccgacaacc ttgattggag acttgaccaa
     6541 acctctggcg aagaagtcca aagctggatc cgcgcgtaat acgactcact atagggcgaa
     6601 ttgggtaccg ggccccccct cgaggtcgac ctgcagcggc catcatgaat taactacacc
     6661 actttgtaca agaaagctgg gtgcggccgc tcatgcatat gcggttttgc aatcgatagc
     6721 tctgaacacc tctactacgt tcatatgatc tcttgtttct ttgggatctt tagaaagtat
     6781 tgtgtgcgct tgtatatgat ggtagtgtgg aatgtcgact ggtccaccgc caagaggagt
     6841 atttaattgt tggtgcgcag caattatttg tcctcctcca attacgtcat gaacaatgtt
     6901 gatgtagcat ttgattccat tattttgtac atctggaaga acatagatgc aatgagggtt
     6961 gtagttaaat tccaattttt tgcccaagat gttaccgcct tttttaaatc ctgaaccgtt
     7021 caacgtaact cggttataaa ctgatccttt ctcgtacgtg acttcagcgc gtgtcttata
     7081 aacgccgtcg ttttcaaaag tgattgttcg ctcttgaacg taaccttctg gcatagcact
     7141 ttttgggtag tctttgatat catttggata ttttgcaaaa cactgtgcac cgtatgtgac
     7201 agttgttaga atgcttgacc aaggtactgg tagttcacca gttgtgcaga tgaaactggc
     7261 ttcaatttta ccagtgttag catcaccatg tcctttacct cttactgaaa acttcattcc
     7321 ctcaacatca ccttccatct caacaacata ggggattttt tctctgaaca aaagtgctcc
     7381 gtaactcatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact gcagaattcg
     7441 aagcttccct cccccatagc tctgtttcca gagtgaaagg agaagcaaac aggagagaca
     7501 gagccccagc accaccacag gatccatggg agccttctct tcctgacaac tggaagctgc
     7561 aaggggtacc agatctgagt ccggtagcgc tagcagcctg cttttttgta caaacttgtg
     7621 atggccgcga attcactagt cgaggctggg gtatattttt tttctttctt ttgaatgtgt
     7681 gtataaaaga gagaaaaaat ggttattttc caattaatta attattaacg attgttaaag
     7741 aaaaaattta tcaacgaggt tgatagaaaa aaaaagtttt tgtagatata gaagaaaaaa
     7801 agattttcac tattggtgtt ataggaaaga aagagaaaaa atttaaga
//
