LOCUS       sequence-332103-        7688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses yotagRFPT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7688)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 7688)
  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: yotagRFPT pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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     168..190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(233..688)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(370..391)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     582..601
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1496..1520
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1525..1534
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2274..2298)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2360..2379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2361..2379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2527..2714
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2582..2606)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2726..2743)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2726..2742)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2735..2757)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3178..4524
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4635..5438)
                     /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(5439..5654)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5679..5697)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5765..5869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5870..5938,5939..6730)
                     /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(6088..6107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6901..7489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7390..7409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7643..7660
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagct taccgcatca ggaaattgta
      241 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
      301 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
      361 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
      421 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
      481 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
      541 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
      601 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
      661 gcgcttaatg cgccgctaca gggcgcgtcc attcgccaag cttcctgaaa cggagaaaca
      721 taaacaggca ttgctgggat cacccataca tcactctgtt ttgcctgacc ttttccggta
      781 atttgaaaac aaacccggtc tcgaagcgga gatccggcga taattaccgc agaaataaac
      841 ccatacacga gacgtagaac cagccgcaca tggccggaga aactcctgcg agaatttcgt
      901 aaactcgcgc gcattgcatc tgtatttcct aatgcggcac ttccaggcct cgatcgagac
      961 cgtttatcca ttgctttttt gttgtctttt tccctcgttc acagaaagtc tgaagaagct
     1021 atagtagaac tatgagcttt ttttgtttct gttttccttt tttttttttt tacctctgtg
     1081 gaaattgtta ctctcacact ctttagttcg tttgtttgtt ttgtttattc caattatgac
     1141 cggtgacgaa acgtggtcga tggtgggtac cgcttatgct cccctccatt agtttcgatt
     1201 atataaaaag gccaaatatt gtattatttt caaatgtcct atcattatcg tctaacatct
     1261 aatttctctt aaattttttc tctttctttc ctataacacc aatagtgaaa atcttttttt
     1321 cttctatatc tacaaaaact ttttttttct atcaacctcg ttgataaatt ttttctttaa
     1381 caatcgttaa taattaatta attggaaaat aaccattttt tctctctttt atacacacat
     1441 tcaaaagaaa gaaaaaaaat ataccccagc ctcgactagt gaattcgcgg ccatcacaag
     1501 tttgtacaaa aaagcaggct gctagccacc atggtatcta aaggtgaaga gttgataaag
     1561 gaaaatatgc acatgaaact ctatatggaa ggcacagtga ataaccatca ttttaagtgt
     1621 acttcagaag gagaaggtaa gccttatgag gggacacaaa ctatgagaat taaagttgtt
     1681 gaaggaggtc cactaccatt cgcttttgac atcctggcaa cgagcttcat gtatggctcc
     1741 agaacattca taaaccatac acaaggtatt ccagatttct tcaaacaaag ttttccagaa
     1801 ggctttactt gggagagagt tactacctac gaggacgggg gtgttttaac tgcgacacag
     1861 gatacatcat tacaagatgg ctgtttgatc tacaacgtaa aaatcagagg tgtcaatttt
     1921 ccatctaacg gacctgttat gcagaaaaag accttgggtt gggaggctaa tactgaaatg
     1981 ctatacccag cagatggagg attggaaggt aggacagata tggcattgaa acttgtagga
     2041 ggcggtcacc taatttgcaa ctttaagaca acctacagaa gcaaaaagcc tgctaaaaat
     2101 cttaaaatgc caggggtgta ctacgtggat catagacttg aacgtatcaa ggaagctgac
     2161 aaagagactt acgttgaaca acacgaagtc gccgtcgcca gatactgtga cttaccttca
     2221 aagttaggtc ataaactgaa tggtatggat gaattgtata agtaagcggc cgcacccagc
     2281 tttcttgtac aaagtggtgt agttaattca tgatggccgc tgcaggtcga cctcgagggg
     2341 gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcggatcc agctttggac
     2401 ttcttcgcca gaggtttggt caagtctcca atcaaggttg tcggcttgtc taccttgcca
     2461 gaaatttacg aaaagatgga aaagggtcaa atcgttggta gatacgttgt tgacacttct
     2521 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     2581 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     2641 aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac
     2701 cacacctcta ccggcatgcc aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2761 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2821 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2881 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataatcg
     2941 gatcgtactt gttacccatc attgaatttt gaacatccga acctgggagt tttccctgaa
     3001 acagatagta tatttgaacc tgtataataa tatatagtct agcgctttac ggaagacaat
     3061 gtatgtattt cggttcctgg agaaactatt gcatctattg cataggtaat cttgcacgtc
     3121 gcatccccgg ttcattttct gcgtttccat cttgcacttc aatagcatat ctttgttaac
     3181 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     3241 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     3301 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     3361 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     3421 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     3481 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     3541 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     3601 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     3661 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     3721 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     3781 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     3841 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     3901 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     3961 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     4021 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     4081 tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt ttggttttca aaagcgctct
     4141 gaagttccta tactttctag ctagagaata ggaacttcgg aataggaact tcaaagcgtt
     4201 tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca
     4261 cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt
     4321 gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc
     4381 tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact
     4441 accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc
     4501 tatcatttcc tttgatattg gatcgatccg atgataagct gtcaaacatg agaattgggt
     4561 aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact
     4621 tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct
     4681 tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca
     4741 acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc
     4801 aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct
     4861 tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc
     4921 ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac
     4981 aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc
     5041 aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct
     5101 gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat
     5161 tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact
     5221 gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg
     5281 ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca
     5341 cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga
     5401 tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcat
     5461 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
     5521 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     5581 gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa
     5641 aatgatgaat tgaaaagcta attcttgaag acgaaagggc ctcgtgatac gcctattttt
     5701 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5761 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5821 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5881 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5941 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6001 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6061 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6121 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6181 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6241 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6301 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6361 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6421 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6481 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6541 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6601 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6661 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6721 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6841 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6901 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6961 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7021 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7081 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7141 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7201 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7261 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7321 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7381 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7441 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7501 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7561 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7621 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     7681 acgcaaac
//
