LOCUS       sequence-332004-        7856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-tagRFPT to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7856)
  AUTHORS   Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B
  TITLE     In vivo characterisation of fluorescent proteins in budding yeast.
  JOURNAL   Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z.
  PUBMED    30783202
REFERENCE   2  (bases 1 to 7856)
  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: "Sci Rep."; 
            date: "2019-02-19"; volume: "9(1):2234. doi"; pages: " 
            10.1038/s41598-019-38913-z"
COMMENT     Sequence Label: cyterm-tagRFPT pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    196..220
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      420..429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1142..1166)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1228..1247)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1229..1247)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1395..1582
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1450..1474)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1594..1611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1594..1610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1603..1625)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2046..3392
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3503..4306)
                     /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(4307..4522)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4547..4565)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4633..4737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4738..4806,4807..5598)
                     /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(4956..4975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5769..6357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6258..6277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6511..6528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6645..6666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6681..6686,6687..6704,6705..6711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6719..6735
                     /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     6724..6746
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6743..6759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6743..6759
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(6789..7244)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6926..6947)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7138..7157
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aatagtgaaa atcttttttt cttctatatc tacaaaaact ttttttttct atcaacctcg
       61 ttgataaatt ttttctttaa caatcgttaa taattaatta attggaaaat aaccattttt
      121 tctctctttt atacacacat tcaaaagaaa gaaaaaaaat ataccccagc ctcgactagt
      181 gaattcgcgg ccatcacaag tttgtacaaa aaagcaggct gctagcgcta ccggactcag
      241 atctggtacc ccttgcagct tccagttgtc aggaagagaa ggctcccatg gatcctgtgg
      301 tggtgctggg gctctgtctc tcctgtttgc ttctcctttc actctggaaa cagagctatg
      361 ggggagggaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg
      421 ccaccatggt gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctctaca
      481 tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct
      541 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
      601 tcgacatcct ggctaccagc ttcatgtacg gcagcagaac cttcatcaac cacacccagg
      661 gcatccccga cttctttaag cagtccttcc ctgagggctt cacatgggag agagtcacca
      721 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
      781 tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct gtgatgcaga
      841 agaaaacact cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctgg
      901 aaggcagaac cgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaacttca
      961 agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg
     1021 tggaccacag actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg
     1081 aggtggctgt ggccagatac tgcgacctcc ctagcaaact gctgtacaag taagcggccg
     1141 cacccagctt tcttgtacaa agtggtgtag ttaattcatg atggccgctg caggtcgacc
     1201 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcggatccag
     1261 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     1321 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     1381 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     1441 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     1501 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     1561 ttattgacca cacctctacc ggcatgccaa ttcactggcc gtcgttttac aacgtcgtga
     1621 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1681 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1741 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     1801 cataatcgga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     1861 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     1921 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     1981 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     2041 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     2101 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     2161 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     2221 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     2281 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     2341 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     2401 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     2461 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     2521 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     2581 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     2641 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     2701 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     2761 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     2821 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     2881 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     2941 cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     3001 agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa taggaacttc
     3061 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     3121 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     3181 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     3241 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     3301 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     3361 ttcaatgcta tcatttcctt tgatattgga tcgatccgat gataagctgt caaacatgag
     3421 aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     3481 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     3541 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     3601 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     3661 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     3721 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     3781 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     3841 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     3901 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     3961 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     4021 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     4081 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     4141 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     4201 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     4261 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     4321 tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     4381 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     4441 tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat
     4501 aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac gaaagggcct cgtgatacgc
     4561 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4621 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4681 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4741 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4801 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4861 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4921 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4981 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5041 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5101 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5161 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5221 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5281 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5341 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5401 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5461 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5521 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5581 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5641 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5701 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5761 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5821 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5881 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5941 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6001 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6061 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6121 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6181 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6241 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6301 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6361 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6421 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6481 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6541 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6601 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6661 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6721 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttacc
     6781 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6841 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6901 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6961 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     7021 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     7081 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     7141 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     7201 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccaagcttc
     7261 ctgaaacgga gaaacataaa caggcattgc tgggatcacc catacatcac tctgttttgc
     7321 ctgacctttt ccggtaattt gaaaacaaac ccggtctcga agcggagatc cggcgataat
     7381 taccgcagaa ataaacccat acacgagacg tagaaccagc cgcacatggc cggagaaact
     7441 cctgcgagaa tttcgtaaac tcgcgcgcat tgcatctgta tttcctaatg cggcacttcc
     7501 aggcctcgat cgagaccgtt tatccattgc ttttttgttg tctttttccc tcgttcacag
     7561 aaagtctgaa gaagctatag tagaactatg agcttttttt gtttctgttt tccttttttt
     7621 tttttttacc tctgtggaaa ttgttactct cacactcttt agttcgtttg tttgttttgt
     7681 ttattccaat tatgaccggt gacgaaacgt ggtcgatggt gggtaccgct tatgctcccc
     7741 tccattagtt tcgattatat aaaaaggcca aatattgtat tattttcaaa tgtcctatca
     7801 ttatcgtcta acatctaatt tctcttaaat tttttctctt tctttcctat aacacc
//
