LOCUS       sequence-326063-        7884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-yotagRFPT to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7884)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 7884)
  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: Cyterm-yotagRFPT  pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    801..825
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1025..1034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1774..1798)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1860..1879)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1861..1879)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2027..2214
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2082..2106)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2226..2243)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2226..2242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2235..2257)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2678..4024
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4135..4938)
                     /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(4939..5154)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5179..5197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5265..5369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5370..5438,5439..6230)
                     /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(5588..5607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6401..6989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6890..6909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7143..7160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7277..7298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7313..7318,7319..7336,7337..7343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7351..7367
                     /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     7356..7378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7375..7391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7375..7391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7421..7876)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7558..7579)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7770..7789
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgaa
      781 ctagtgaatt cgcggccatc acaagtttgt acaaaaaagc aggctgctag cgctaccgga
      841 ctcagatctg gtaccccttg cagcttccag ttgtcaggaa gagaaggctc ccatggatcc
      901 tgtggtggtg ctggggctct gtctctcctg tttgcttctc ctttcactct ggaaacagag
      961 ctatggggga gggaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc
     1021 ggtcgccacc atggtatcta aaggtgaaga gttgataaag gaaaatatgc acatgaaact
     1081 ctatatggaa ggcacagtga ataaccatca ttttaagtgt acttcagaag gagaaggtaa
     1141 gccttatgag gggacacaaa ctatgagaat taaagttgtt gaaggaggtc cactaccatt
     1201 cgcttttgac atcctggcaa cgagcttcat gtatggctcc agaacattca taaaccatac
     1261 acaaggtatt ccagatttct tcaaacaaag ttttccagaa ggctttactt gggagagagt
     1321 tactacctac gaggacgggg gtgttttaac tgcgacacag gatacatcat tacaagatgg
     1381 ctgtttgatc tacaacgtaa aaatcagagg tgtcaatttt ccatctaacg gacctgttat
     1441 gcagaaaaag accttgggtt gggaggctaa tactgaaatg ctatacccag cagatggagg
     1501 attggaaggt aggacagata tggcattgaa acttgtagga ggcggtcacc taatttgcaa
     1561 ctttaagaca acctacagaa gcaaaaagcc tgctaaaaat cttaaaatgc caggggtgta
     1621 ctacgtggat catagacttg aacgtatcaa ggaagctgac aaagagactt acgttgaaca
     1681 acacgaagtc gccgtcgcca gatactgtga cttaccttca aagttaggtc ataaactgaa
     1741 tggtatggat gaattgtata agtaagcggc cgcacccagc tttcttgtac aaagtggtgt
     1801 agttaattca tgatggccgc tgcaggtcga cctcgagggg gggcccggta cccaattcgc
     1861 cctatagtga gtcgtattac gcgcggatcc agctttggac ttcttcgcca gaggtttggt
     1921 caagtctcca atcaaggttg tcggcttgtc taccttgcca gaaatttacg aaaagatgga
     1981 aaagggtcaa atcgttggta gatacgttgt tgacacttct aaataagcga atttcttatg
     2041 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     2101 tgactcttag gttttaaaac ganaattctt attcttgagt aactctttcc tgtaggtcag
     2161 gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgcc
     2221 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2281 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2341 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     2401 ctccttacgc atctgtgcgg tatttcacac cgcataatcg gatcgtactt gttacccatc
     2461 attgaatttt gaacatccga acctgggagt tttccctgaa acagatagta tatttgaacc
     2521 tgtataataa tatatagtct agcgctttac ggaagacaat gtatgtattt cggttcctgg
     2581 agaaactatt gcatctattg cataggtaat cttgcacgtc gcatccccgg ttcattttct
     2641 gcgtttccat cttgcacttc aatagcatat ctttgttaac gaagcatctg tgcttcattt
     2701 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     2761 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     2821 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     2881 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     2941 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     3001 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     3061 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     3121 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     3181 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     3241 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     3301 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     3361 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     3421 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     3481 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     3541 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
     3601 agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
     3661 ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg
     3721 aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt
     3781 gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc
     3841 gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta
     3901 tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat
     3961 gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg
     4021 gatcgatccg atgataagct gtcaaacatg agaattgggt aataactgat ataattaaat
     4081 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     4141 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     4201 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     4261 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     4321 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     4381 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     4441 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     4501 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     4561 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     4621 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     4681 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     4741 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     4801 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     4861 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     4921 atatgtagct ttcgacatga tttatcttcg tttcctgcat gtttttgttc tgtgcagttg
     4981 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     5041 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     5101 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagcta
     5161 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5221 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5281 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5341 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5401 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5461 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5521 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5581 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5641 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5701 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5761 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5821 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5881 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5941 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6001 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6061 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6121 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6181 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6241 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7141 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7201 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     7261 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     7321 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     7381 acagctatga ccatgattac gccaagctta ccgcatcagg aaattgtaag cgttaatatt
     7441 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7501 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     7561 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7621 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     7681 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7741 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     7801 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7861 ccgctacagg gcgcgtccat tcgc
//
