LOCUS       sequence-331864-        8456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces equimolar expression of mAvicFP1 and mCherry for use in 
            yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8456)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8456)
  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"
COMMENT     Sequence Label: mAvicFP1-T2A-mCherry pdrf1-GW
FEATURES             Location/Qualifiers
     source          1..8456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1976..1994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2019..2234
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2235..3038
                     /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      3149..4495
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     4916..4938
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4930..4947
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4931..4947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(4959..5146)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5067..5091
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     5294..5313
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5294..5312
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5375..5399
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5409..6119)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5687..5706)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5956..5974
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6116..6125
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6141..6194)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(6921..6945)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      7753..8208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7840..7859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8050..8071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8238..8254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8238..8254)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8251..8273)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8262..8278
                     /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(8286..8292,8293..8310,8311..8316))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8331..8352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1981 gcgtatcacg aggccctttc gtcttcaaga attagctttt caattcatca tttttttttt
     2041 attctttttt ttgatttcgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     2101 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     2161 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acatgcagga
     2221 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     2281 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     2341 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     2401 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     2461 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg
     2521 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     2581 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     2641 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     2701 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     2761 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     2821 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac
     2881 agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag
     2941 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     3001 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     3061 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccaatt
     3121 ctcatgtttg acagcttatc atcggatcga tccaatatca aaggaaatga tagcattgaa
     3181 ggatgagact aatccaattg aggagtggca gcatatagaa cagctaaagg gtagtgctga
     3241 aggaagcata cgataccccg catggaatgg gataatatca caggaggtac tagactacct
     3301 ttcatcctac ataaatagac gcatataagt acgcatttaa gcataaacac gcactatgcc
     3361 gttcttctca tgtatatata tatacaggca acacgcagat ataggtgcga cgtgaacagt
     3421 gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt
     3481 gaagttccta ttccgaagtt cctattctct agctagaaag tataggaact tcagagcgct
     3541 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
     3601 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     3661 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     3721 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     3781 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     3841 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     3901 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     3961 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     4021 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     4081 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     4141 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     4201 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     4261 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     4321 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     4381 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     4441 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttaacaa
     4501 agatatgcta ttgaagtgca agatggaaac gcagaaaatg aaccggggat gcgacgtgca
     4561 agattaccta tgcaatagat gcaatagttt ctccaggaac cgaaatacat acattgtctt
     4621 ccgtaaagcg ctagactata tattattata caggttcaaa tatactatct gtttcaggga
     4681 aaactcccag gttcggatgt tcaaaattca atgatgggta acaagtacga tccgattatg
     4741 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     4801 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4861 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     4921 tcacgacgtt gtaaaacgac ggccagtgaa ttggcatgcc ggtagaggtg tggtcaataa
     4981 gagcgacctc atgctatacc tgagaaagca acctgaccta caggaaagag ttactcaaga
     5041 ataagaattt tcgttttaaa acctaagagt cactttaaaa tttgtataca cttatttttt
     5101 ttataactta tttaataata aaaatcataa atcataagaa attcgcttat ttagaagtgt
     5161 caacaacgta tctaccaacg atttgaccct tttccatctt ttcgtaaatt tttggcaagg
     5221 tagacaagcc gacaaccttg attggagact tgaccaaacc tctggcgaag aagtccaaag
     5281 ctggatccgc gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga
     5341 ggtcgacctg cagcggccat catgaattaa ctacaccact ttgtacaaga aagctgggtg
     5401 cggccgcttt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
     5461 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
     5521 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     5581 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     5641 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     5701 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     5761 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     5821 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     5881 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     5941 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     6001 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     6061 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg
     6121 gtggcgaccg gtggatcccc agggccggga ttctcctcca cgtcaccgca tgttagaaga
     6181 cttcctctgc cctctccgga cttgtacagt tcgtccatac catgggtgat accgcaagcg
     6241 gtaacgaatt caaccaggtt catatggtca cgcttttcgt tagggtcctt ggtgatcttg
     6301 gtctgggtag acaggtaatg gtagtcaggc agcagaacag gaccgtcacc gatagggatg
     6361 ttctgctggt catggtcagc cagctggata ccttcaccct tcaggttatg acggatcttg
     6421 aagttaacct tgataccgtt gttagccttg tcagacagaa cgtaaacaga atgagagttg
     6481 tagttgtatt ccatcttgtt acccaggatg ttaccgtctt ccttgaagtc gatacccttc
     6541 agttcgatac ggttaaccag ggtgtcacct tcgaacttaa cttcagcacg ggtcttgtaa
     6601 tgaccgtcgt cctggaaaga gatggtacgc ttctggatgt aaccgtcagg catagcagac
     6661 ttgtagaagt catgctgctt catatgttca gggtaacggg tgaagcactg aacaccgtag
     6721 cataaggtgg taaccagggt aggccaagga acaggcaggg taccggtggt gcaaacgaac
     6781 ttgatttcga tcttaccaga accagcgtca ccttcacctt caccacgaac agagaactta
     6841 tgaccatgaa cgtcaccgtt cagttcgatc aggataggaa cgataccgtt gaacagttca
     6901 gcacccttag acatgctagc agcctgcttt tttgtacaaa cttgtgatgg ccgcgaattc
     6961 actagtcgag gctggggtat attttttttc tttcttttga atgtgtgtat aaaagagaga
     7021 aaaaatggtt attttccaat taattaatta ttaacgattg ttaaagaaaa aatttatcaa
     7081 cgaggttgat agaaaaaaaa agtttttgta gatatagaag aaaaaaagat tttcactatt
     7141 ggtgttatag gaaagaaaga gaaaaaattt aagagaaatt agatgttaga cgataatgat
     7201 aggacatttg aaaataatac aatatttggc ctttttatat aatcgaaact aatggagggg
     7261 agcataagcg gtacccacca tcgaccacgt ttcgtcaccg gtcataattg gaataaacaa
     7321 aacaaacaaa cgaactaaag agtgtgagag taacaatttc cacagaggta aaaaaaaaaa
     7381 aaaggaaaac agaaacaaaa aaagctcata gttctactat agcttcttca gactttctgt
     7441 gaacgaggga aaaagacaac aaaaaagcaa tggataaacg gtctcgatcg aggcctggaa
     7501 gtgccgcatt aggaaataca gatgcaatgc gcgcgagttt acgaaattct cgcaggagtt
     7561 tctccggcca tgtgcggctg gttctacgtc tcgtgtatgg gtttatttct gcggtaatta
     7621 tcgccggatc tccgcttcga gaccgggttt gttttcaaat taccggaaaa ggtcaggcaa
     7681 aacagagtga tgtatgggtg atcccagcaa tgcctgttta tgtttctccg tttcaggaag
     7741 cttggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     7801 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     7861 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     7921 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     7981 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     8041 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     8101 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     8161 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     8221 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     8281 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     8341 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     8401 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcg
//
