LOCUS       sequence-332006-        7865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-mAvicFP1 to check for monomericity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7865)
  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 7865)
  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-mAvicFP1 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(249..704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(386..407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     598..617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1512..1536
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2467..2491)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2553..2572)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2554..2572)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2720..2907
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2775..2799)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2919..2936)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2919..2935)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2928..2950)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3371..4717
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4828..5631)
                     /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(5632..5847)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5872..5890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5958..6062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6063..6131,6132..6923)
                     /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(6281..6300)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7094..7682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7583..7602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7836..7853
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttacc
      241 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
      301 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      361 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
      421 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
      481 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
      541 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
      601 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
      661 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccaagcttc
      721 ctgaaacgga gaaacataaa caggcattgc tgggatcacc catacatcac tctgttttgc
      781 ctgacctttt ccggtaattt gaaaacaaac ccggtctcga agcggagatc cggcgataat
      841 taccgcagaa ataaacccat acacgagacg tagaaccagc cgcacatggc cggagaaact
      901 cctgcgagaa tttcgtaaac tcgcgcgcat tgcatctgta tttcctaatg cggcacttcc
      961 aggcctcgat cgagaccgtt tatccattgc ttttttgttg tctttttccc tcgttcacag
     1021 aaagtctgaa gaagctatag tagaactatg agcttttttt gtttctgttt tccttttttt
     1081 tttttttacc tctgtggaaa ttgttactct cacactcttt agttcgtttg tttgttttgt
     1141 ttattccaat tatgaccggt gacgaaacgt ggtcgatggt gggtaccgct tatgctcccc
     1201 tccattagtt tcgattatat aaaaaggcca aatattgtat tattttcaaa tgtcctatca
     1261 ttatcgtcta acatctaatt tctcttaaat tttttctctt tctttcctat aacaccaata
     1321 gtgaaaatct ttttttcttc tatatctaca aaaacttttt ttttctatca acctcgttga
     1381 taaatttttt ctttaacaat cgttaataat taattaattg gaaaataacc attttttctc
     1441 tcttttatac acacattcaa aagaaagaaa aaaaatatac cccagcctcg actagtgaat
     1501 tcgcggccat cacaagtttg tacaaaaaag caggctgcta gcgctaccgg actcagatct
     1561 ggtacccctt gcagcttcca gttgtcagga agagaaggct cccatggatc ctgtggtggt
     1621 gctggggctc tgtctctcct gtttgcttct cctttcactc tggaaacaga gctatggggg
     1681 agggaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac
     1741 catgtctaag ggtgctgaac tgttcaacgg tatcgttcct atcctgatcg aactgaacgg
     1801 tgacgttcat ggtcataagt tctctgttcg tggtgaaggt gaaggtgacg ctggttctgg
     1861 taagatcgaa atcaagttcg tttgcaccac cggtaccctg cctgttcctt ggcctaccct
     1921 ggttaccacc ttatgctacg gtgttcagtg cttcacccgt taccctgaac atatgaagca
     1981 gcatgacttc tacaagtctg ctatgcctga cggttacatc cagaagcgta ccatctcttt
     2041 ccaggacgac ggtcattaca agacccgtgc tgaagttaag ttcgaaggtg acaccctggt
     2101 taaccgtatc gaactgaagg gtatcgactt caaggaagac ggtaacatcc tgggtaacaa
     2161 gatggaatac aactacaact ctcattctgt ttacgttctg tctgacaagg ctaacaacgg
     2221 tatcaaggtt aacttcaaga tccgtcataa cctgaagggt gaaggtatcc agctggctga
     2281 ccatgaccag cagaacatcc ctatcggtga cggtcctgtt ctgctgcctg actaccatta
     2341 cctgtctacc cagaccaaga tcaccaagga ccctaacgaa aagcgtgacc atatgaacct
     2401 ggttgaattc gttaccgctt gcggtatcac ccatggtatg gacgaactgt acaagtgagc
     2461 ggccgcaccc agctttcttg tacaaagtgg tgtagttaat tcatgatggc cgctgcaggt
     2521 cgacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgga
     2581 tccagctttg gacttcttcg ccagaggttt ggtcaagtct ccaatcaagg ttgtcggctt
     2641 gtctaccttg ccagaaattt acgaaaagat ggaaaagggt caaatcgttg gtagatacgt
     2701 tgttgacact tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt
     2761 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     2821 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt
     2881 cgctcttatt gaccacacct ctaccggcat gccaattcac tggccgtcgt tttacaacgt
     2941 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3001 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3061 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3121 caccgcataa tcggatcgta cttgttaccc atcattgaat tttgaacatc cgaacctggg
     3181 agttttccct gaaacagata gtatatttga acctgtataa taatatatag tctagcgctt
     3241 tacggaagac aatgtatgta tttcggttcc tggagaaact attgcatcta ttgcataggt
     3301 aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac ttcaatagca
     3361 tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     3421 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     3481 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     3541 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3601 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     3661 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     3721 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3781 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3841 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3901 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3961 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     4021 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     4081 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     4141 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     4201 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     4261 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt
     4321 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cggaatagga
     4381 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     4441 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     4501 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     4561 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     4621 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     4681 catccttcaa tgctatcatt tcctttgata ttggatcgat ccgatgataa gctgtcaaac
     4741 atgagaattg ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     4801 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     4861 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     4921 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     4981 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     5041 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     5101 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     5161 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     5221 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     5281 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     5341 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     5401 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     5461 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     5521 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     5581 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     5641 tcgtttcctg catgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     5701 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     5761 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     5821 agaataaaaa aaaaatgatg aattgaaaag ctaattcttg aagacgaaag ggcctcgtga
     5881 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5941 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6001 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6061 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6121 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6181 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6241 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6301 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6361 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6421 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6481 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6541 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6601 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6661 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6721 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6781 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6841 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6901 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6961 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7021 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7201 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7261 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7321 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7381 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7441 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7501 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7561 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7621 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7681 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7741 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7801 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7861 aagag
//
