LOCUS       sequence-332367-        7868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-ymTq2 to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7868)
  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 7868)
  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-ymtq2 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      612..1067
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(699..718)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     909..930
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1097..1113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1097..1113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1110..1132)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1121..1137
                     /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(1145..1151,1152..1169,1170..1175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1190..1211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1328..1345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1499..2087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1579..1598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2258..3049,3050..3118))
                     /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     2881..2900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3119..3223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3291..3309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3334..3549
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3550..4353
                     /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      4464..5810
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6231..6253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6245..6262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6246..6262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(6274..6461)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6382..6406
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     6609..6628
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6609..6627
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6690..6714
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      7439..7448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(7648..7672)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtgttatagg aaagaaagag aaaaaattta agagaaatta gatgttagac gataatgata
       61 ggacatttga aaataataca atatttggcc tttttatata atcgaaacta atggagggga
      121 gcataagcgg tacccaccat cgaccacgtt tcgtcaccgg tcataattgg aataaacaaa
      181 acaaacaaac gaactaaaga gtgtgagagt aacaatttcc acagaggtaa aaaaaaaaaa
      241 aaggaaaaca gaaacaaaaa aagctcatag ttctactata gcttcttcag actttctgtg
      301 aacgagggaa aaagacaaca aaaaagcaat ggataaacgg tctcgatcga ggcctggaag
      361 tgccgcatta ggaaatacag atgcaatgcg cgcgagttta cgaaattctc gcaggagttt
      421 ctccggccat gtgcggctgg ttctacgtct cgtgtatggg tttatttctg cggtaattat
      481 cgccggatct ccgcttcgag accgggtttg ttttcaaatt accggaaaag gtcaggcaaa
      541 acagagtgat gtatgggtga tcccagcaat gcctgtttat gtttctccgt ttcaggaagc
      601 ttggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1021 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtaa
     1081 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1141 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1201 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1261 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1381 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1441 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1501 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1561 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1621 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1681 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1741 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1801 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1861 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1921 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1981 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2041 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2101 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2161 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2221 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2281 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2341 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2401 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2461 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2521 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2581 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2641 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2701 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2761 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2821 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2881 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2941 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3001 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3061 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3121 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3181 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3241 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3301 tcacgaggcc ctttcgtctt caagaattag cttttcaatt catcattttt tttttattct
     3361 tttttttgat ttcggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     3421 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     3481 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacatg caggaaacga
     3541 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     3601 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     3661 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     3721 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     3781 aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt tgctgacatt
     3841 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     3901 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     3961 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     4021 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     4081 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     4141 ttgattatga cacccggtgt gggtttagat gacaagggag acgcattggg tcaacagtat
     4201 agaaccgtgg atgatgtggt ctctacagga tctgacatta ttattgttgg aagaggacta
     4261 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     4321 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     4381 atactaaact cacaaattag agcttcaatt taattatatc agttattacc caattctcat
     4441 gtttgacagc ttatcatcgg atcgatccaa tatcaaagga aatgatagca ttgaaggatg
     4501 agactaatcc aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa
     4561 gcatacgata ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat
     4621 cctacataaa tagacgcata taagtacgca tttaagcata aacacgcact atgccgttct
     4681 tctcatgtat atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct
     4741 gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac gctttgaagt
     4801 tcctattccg aagttcctat tctctagcta gaaagtatag gaacttcaga gcgcttttga
     4861 aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg
     4921 agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta tctctttgct
     4981 atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct ccttgaactt
     5041 gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc tttagacaaa
     5101 aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt
     5161 tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt ctgaccaatg
     5221 aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac atcggtatag
     5281 aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag
     5341 taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca ccaaagtagc
     5401 cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg cattatagag
     5461 cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc gctctcggga
     5521 tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata gcgctctcgc
     5581 gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct
     5641 cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct
     5701 ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     5761 gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt aacaaagata
     5821 tgctattgaa gtgcaagatg gaaacgcaga aaatgaaccg gggatgcgac gtgcaagatt
     5881 acctatgcaa tagatgcaat agtttctcca ggaaccgaaa tacatacatt gtcttccgta
     5941 aagcgctaga ctatatatta ttatacaggt tcaaatatac tatctgtttc agggaaaact
     6001 cccaggttcg gatgttcaaa attcaatgat gggtaacaag tacgatccga ttatgcggtg
     6061 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6241 acgttgtaaa acgacggcca gtgaattggc atgccggtag aggtgtggtc aataagagcg
     6301 acctcatgct atacctgaga aagcaacctg acctacagga aagagttact caagaataag
     6361 aattttcgtt ttaaaaccta agagtcactt taaaatttgt atacacttat tttttttata
     6421 acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga agtgtcaaca
     6481 acgtatctac caacgatttg acccttttcc atcttttcgt aaatttctgg caaggtagac
     6541 aagccgacaa ccttgattgg agacttgacc aaacctctgg cgaagaagtc caaagctgga
     6601 tccgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     6661 acctgcagcg gccatcatga attaactaca ccactttgta caagaaagct gggtgcggcc
     6721 gcttatttat acaattcatc catacctaaa gtgatcccag cagcagtaac gaattccaac
     6781 aaaaccatat gatctctctt ttcgtttgga tccttagaca acttagattg agtagacaag
     6841 taatggttat ctggcaacaa aactggaccg tcaccaattg gagtgttttg ttggtaatga
     6901 tcagctaatt gaacaccacc atcttcaatg ttatgtctaa tcttgaagtt cgccttaata
     6961 ccgttctttt gcttatcagc agtaatgtaa acgttatcag agaagtaatt atattccaac
     7021 ttatgaccca aaatattacc atcttcctta aaatcaatac ccttcaattc aattctatta
     7081 actaaagtat ctccttcgaa cttaacttca gctctagtct tatagttacc atcatccttg
     7141 aagaagatag ttctttcttg aacgtaacct tctggcatag cagacttaaa gaaatcatgt
     7201 tgcttcatat gatctgggta tctagcgaaa cattgaacac cccaagacaa agtagtaacc
     7261 aaagttggcc atggaactgg taatttacct gtagtacaaa tgaatttcaa agttaattta
     7321 ccgtatgttg cgtcaccttc accttcaccg cttacagaga acttatgacc gttaacgtca
     7381 ccatctaatt ctactaaaat tggcactaca ccagtgaaca attcttcacc tttactaacc
     7441 atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcgaagcttc
     7501 cctcccccat agctctgttt ccagagtgaa aggagaagca aacaggagag acagagcccc
     7561 agcaccacca caggatccat gggagccttc tcttcctgac aactggaagc tgcaaggggt
     7621 accagatctg agtccggtag cgctagcagc ctgctttttt gtacaaactt gtgatggccg
     7681 cgaattcact agtcgaggct ggggtatatt ttttttcttt cttttgaatg tgtgtataaa
     7741 agagagaaaa aatggttatt ttccaattaa ttaattatta acgattgtta aagaaaaaat
     7801 ttatcaacga ggttgataga aaaaaaaagt ttttgtagat atagaagaaa aaaagatttt
     7861 cactattg
//
