LOCUS       sequence-332003-        7883 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 7883)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 7883)
  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..7883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(232..687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(369..390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     581..600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1495..1519
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1719..1728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2468..2492)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2554..2573)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2555..2573)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2721..2908
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2776..2800)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2920..2937)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2920..2936)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2929..2951)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3372..4718
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4829..5632)
                     /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(5633..5848)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5873..5891)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5959..6063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6064..6132,6133..6924)
                     /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(6282..6301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7095..7683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7584..7603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7837..7854
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagctt accgcatcag gaaattgtaa
      241 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
      301 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
      361 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
      421 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
      481 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
      541 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
      601 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
      661 cgcttaatgc gccgctacag ggcgcgtcca ttcgccaagc ttcctgaaac ggagaaacat
      721 aaacaggcat tgctgggatc acccatacat cactctgttt tgcctgacct tttccggtaa
      781 tttgaaaaca aacccggtct cgaagcggag atccggcgat aattaccgca gaaataaacc
      841 catacacgag acgtagaacc agccgcacat ggccggagaa actcctgcga gaatttcgta
      901 aactcgcgcg cattgcatct gtatttccta atgcggcact tccaggcctc gatcgagacc
      961 gtttatccat tgcttttttg ttgtcttttt ccctcgttca cagaaagtct gaagaagcta
     1021 tagtagaact atgagctttt tttgtttctg ttttcctttt tttttttttt acctctgtgg
     1081 aaattgttac tctcacactc tttagttcgt ttgtttgttt tgtttattcc aattatgacc
     1141 ggtgacgaaa cgtggtcgat ggtgggtacc gcttatgctc ccctccatta gtttcgatta
     1201 tataaaaagg ccaaatattg tattattttc aaatgtccta tcattatcgt ctaacatcta
     1261 atttctctta aattttttct ctttctttcc tataacacca atagtgaaaa tctttttttc
     1321 ttctatatct acaaaaactt tttttttcta tcaacctcgt tgataaattt tttctttaac
     1381 aatcgttaat aattaattaa ttggaaaata accatttttt ctctctttta tacacacatt
     1441 caaaagaaag aaaaaaaata taccccagcc tcgactagtg aattcgcggc catcacaagt
     1501 ttgtacaaaa aagcaggctg ctagcgctac cggactcaga tctggtaccc cttgcagctt
     1561 ccagttgtca ggaagagaag gctcccatgg atcctgtggt ggtgctgggg ctctgtctct
     1621 cctgtttgct tctcctttca ctctggaaac agagctatgg gggagggaag cttcgaattc
     1681 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggta tctaaaggtg
     1741 aagagttgat aaaggaaaat atgcacatga aactctatat ggaaggcaca gtgaataacc
     1801 atcattttaa gtgtacttca gaaggagaag gtaagcctta tgaggggaca caaactatga
     1861 gaattaaagt tgttgaagga ggtccactac cattcgcttt tgacatcctg gcaacgagct
     1921 tcatgtatgg ctccagaaca ttcataaacc atacacaagg tattccagat ttcttcaaac
     1981 aaagttttcc agaaggcttt acttgggaga gagttactac ctacgaggac gggggtgttt
     2041 taactgcgac acaggataca tcattacaag atggctgttt gatctacaac gtaaaaatca
     2101 gaggtgtcaa ttttccatct aacggacctg ttatgcagaa aaagaccttg ggttgggagg
     2161 ctaatactga aatgctatac ccagcagatg gaggattgga aggtaggaca gatatggcat
     2221 tgaaacttgt aggaggcggt cacctaattt gcaactttaa gacaacctac agaagcaaaa
     2281 agcctgctaa aaatcttaaa atgccagggg tgtactacgt ggatcataga cttgaacgta
     2341 tcaaggaagc tgacaaagag acttacgttg aacaacacga agtcgccgtc gccagatact
     2401 gtgacttacc ttcaaagtta ggtcataaac tgaatggtat ggatgaattg tataagtaag
     2461 cggccgcacc cagctttctt gtacaaagtg gtgtagttaa ttcatgatgg ccgctgcagg
     2521 tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgg
     2581 atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct
     2641 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg
     2701 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     2761 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     2821 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     2881 tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg
     2941 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3001 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3061 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3121 acaccgcata atcggatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg
     3181 gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct
     3241 ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg
     3301 taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc
     3361 atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     3421 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     3481 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     3541 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     3601 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     3661 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     3721 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     3781 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     3841 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     3901 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     3961 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     4021 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     4081 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     4141 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     4201 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     4261 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt
     4321 ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact tcggaatagg
     4381 aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata
     4441 cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag
     4501 aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag
     4561 gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg
     4621 cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc
     4681 tcatccttca atgctatcat ttcctttgat attggatcga tccgatgata agctgtcaaa
     4741 catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     4801 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     4861 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     4921 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     4981 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     5041 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     5101 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     5161 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     5221 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     5281 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     5341 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     5401 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     5461 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     5521 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     5581 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     5641 ttcgtttcct gcatgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     5701 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     5761 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     5821 aagaataaaa aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa gggcctcgtg
     5881 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5941 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6001 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6061 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6121 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6181 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6241 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6301 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6361 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6421 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6481 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6541 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6601 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6661 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6721 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6781 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6841 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6901 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6961 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7021 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7081 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7141 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7201 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7261 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7321 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7381 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7441 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7501 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7561 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7621 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7681 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7741 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7801 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7861 gaagagcgcc caatacgcaa acc
//
