LOCUS       sequence-332100-        7847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-ymScarletI to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7847)
  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 7847)
  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-ymScarletI pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1264..1288
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1488..1497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2201..2225)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2287..2306)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2288..2306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2454..2641
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2509..2533)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2653..2670)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2653..2669)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2662..2684)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3105..4451
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4562..5365)
                     /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(5366..5581)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5606..5624)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5692..5796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5797..5865,5866..6657)
                     /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(6015..6034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6828..7416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7317..7336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7570..7587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7704..7725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7740..7745,7746..7763,7764..7770)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7778..7794
                     /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     7783..7805
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7802..7818
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7802..7818
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccaagct tcctgaaacg
      481 gagaaacata aacaggcatt gctgggatca cccatacatc actctgtttt gcctgacctt
      541 ttccggtaat ttgaaaacaa acccggtctc gaagcggaga tccggcgata attaccgcag
      601 aaataaaccc atacacgaga cgtagaacca gccgcacatg gccggagaaa ctcctgcgag
      661 aatttcgtaa actcgcgcgc attgcatctg tatttcctaa tgcggcactt ccaggcctcg
      721 atcgagaccg tttatccatt gcttttttgt tgtctttttc cctcgttcac agaaagtctg
      781 aagaagctat agtagaacta tgagcttttt ttgtttctgt tttccttttt ttttttttta
      841 cctctgtgga aattgttact ctcacactct ttagttcgtt tgtttgtttt gtttattcca
      901 attatgaccg gtgacgaaac gtggtcgatg gtgggtaccg cttatgctcc cctccattag
      961 tttcgattat ataaaaaggc caaatattgt attattttca aatgtcctat cattatcgtc
     1021 taacatctaa tttctcttaa attttttctc tttctttcct ataacaccaa tagtgaaaat
     1081 ctttttttct tctatatcta caaaaacttt ttttttctat caacctcgtt gataaatttt
     1141 ttctttaaca atcgttaata attaattaat tggaaaataa ccattttttc tctcttttat
     1201 acacacattc aaaagaaaga aaaaaaatat accccagcct cgactagtga attcgcggcc
     1261 atcacaagtt tgtacaaaaa agcaggctgc tagcgctacc ggactcagat ctggtacccc
     1321 ttgcagcttc cagttgtcag gaagagaagg ctcccatgga tcctgtggtg gtgctggggc
     1381 tctgtctctc ctgtttgctt ctcctttcac tctggaaaca gagctatggg ggagggaagc
     1441 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtta
     1501 gtaaaggtga agctgttata aaagaattca tgaggtttaa agttcatatg gaaggttcaa
     1561 tgaatggtca tgaatttgaa attgaaggtg aaggtgaagg tagaccatat gaaggtacac
     1621 aaactgctaa attgaaagtt actaaaggtg gtccattacc attttcttgg gatattttgt
     1681 ctccacaatt catgtatggt tctagagctt ttattaagca tccagctgat attccagatt
     1741 attataaaca atcttttcct gaaggtttta aatgggaaag agttatgaat ttcgaagatg
     1801 gtggtgctgt tactgttact caagatactt ctttggaaga tggtacttta atctataaag
     1861 ttaaattgag aggtactaat tttccaccag atggtccagt tatgcaaaag aaaactatgg
     1921 gttgggaagc atctactgaa agattgtatc cagaagatgg tgttttgaaa ggagacatta
     1981 agatggcttt gagattgaaa gatggtggtc ggtacctggc cgacttcaag accacctata
     2041 aagctaaaaa accagttcaa atgccaggtg catataatgt tgatagaaag ttagatataa
     2101 cgtcgcataa cgaggactat actgtagttg aacaatatga acgtagtgaa ggtagacata
     2161 gtaccggagg aatggatgaa ttgtataaat aagcggccgc acccagcttt cttgtacaaa
     2221 gtggtgtagt taattcatga tggccgctgc aggtcgacct cgaggggggg cccggtaccc
     2281 aattcgccct atagtgagtc gtattacgcg cggatccagc tttggacttc ttcgccagag
     2341 gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac cttgccagaa atttacgaaa
     2401 agatggaaaa gggtcaaatc gttggtagat acgttgttga cacttctaaa taagcgaatt
     2461 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     2521 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     2581 aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg
     2641 gcatgccaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2701 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2761 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     2821 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataatcggat cgtacttgtt
     2881 acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat
     2941 ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg
     3001 ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc
     3061 attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc
     3121 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     3181 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     3241 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     3301 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     3361 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     3421 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     3481 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     3541 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     3601 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     3661 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     3721 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     3781 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     3841 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     3901 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     3961 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     4021 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     4081 tttctagcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     4141 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     4201 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     4261 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     4321 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     4381 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     4441 gatattggat cgatccgatg ataagctgtc aaacatgaga attgggtaat aactgatata
     4501 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     4561 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     4621 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     4681 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     4741 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     4801 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     4861 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     4921 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     4981 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     5041 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     5101 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     5161 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     5221 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     5281 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     5341 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt
     5401 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     5461 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     5521 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     5581 aaagctaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5641 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5701 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5761 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5821 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5881 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5941 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6001 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6061 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6121 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6181 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6241 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6301 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6361 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6421 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6481 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6541 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6601 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6661 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6721 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6781 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6841 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6901 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6961 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7021 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7081 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7141 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7201 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7261 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7321 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7381 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7441 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7501 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7561 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7621 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7681 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7741 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7801 acaggaaaca gctatgacca tgattacgcc aagcttaccg catcagg
//
