LOCUS       sequence-331869-        7648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AausFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7648)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 7648)
  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: AausFP1 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    232..256
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(970..994)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1056..1075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1057..1075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1223..1410
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1278..1302)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1422..1439)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1422..1438)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1431..1453)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1874..3220
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3331..4134)
                     /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(4135..4350)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4375..4393)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4461..4565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4566..4634,4635..5426)
                     /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(4784..4803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5597..6185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6086..6105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6339..6356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6473..6494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6509..6514,6515..6532,6533..6539)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6547..6563
                     /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     6552..6574
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6571..6587
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6571..6587
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(6617..7072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6754..6775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6966..6985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tctcttaaat tttttctctt tctttcctat aacaccaata gtgaaaatct ttttttcttc
       61 tatatctaca aaaacttttt ttttctatca acctcgttga taaatttttt ctttaacaat
      121 cgttaataat taattaattg gaaaataacc attttttctc tcttttatac acacattcaa
      181 aagaaagaaa aaaaatatac cccagcctcg actagtgaat tcgcggccat cacaagtttg
      241 tacaaaaaag caggctgcta gcatgagtta cggagcactt ttgttcagag aaaaaatccc
      301 ctatgttgtt gagatggaag gtgatgttga gggaatgaag ttttcagtaa gaggtaaagg
      361 acatggtgat gctaacactg gtaaaattga agccagtttc atctgcacaa ctggtgaact
      421 accagtacct tggtcaagca ttctaacaac tgtcacatac ggtgcacagt gttttgcaaa
      481 atatccaaat gatatcaaag actacccaaa aagtgctatg ccagaaggtt acgttcaaga
      541 gcgaacaatc acttttgaaa acgacggcgt ttataagaca cgcgctgaag tcacgtacga
      601 gaaaggatca gtttataacc gagttacgtt gaacggttca ggatttaaaa aaggcggtaa
      661 catcttgggc aaaaaattgg aatttaacta caaccctcat tgcatctatg ttcttccaga
      721 tgtacaaaat aatggaatca aatgctacat caacattgtt catgacgtaa ttggaggagg
      781 acaaataatt gctgcgcacc aacaattaaa tactcctctt ggcggtggac cagtcgacat
      841 tccacactac catcatatac aagcgcacac aatactttct aaagatccca aagaaacaag
      901 agatcatatg aacgtagtag aggtgttcag agctatcgat tgcaaaaccg catatgcata
      961 agcggccgca cccagctttc ttgtacaaag tggtgtagtt aattcatgat ggccgctgca
     1021 ggtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     1081 ggatccagct ttggacttct tcgccagagg tttggtcaag tctccaatca aggttgtcgg
     1141 cttgtctacc ttgccagaaa tttacgaaaa gatggaaaag ggtcaaatcg ttggtagata
     1201 cgttgttgac acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata
     1261 agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa
     1321 attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga
     1381 ggtcgctctt attgaccaca cctctaccgg catgccaatt cactggccgt cgttttacaa
     1441 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1501 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1561 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     1621 tcacaccgca taatcggatc gtacttgtta cccatcattg aattttgaac atccgaacct
     1681 gggagttttc cctgaaacag atagtatatt tgaacctgta taataatata tagtctagcg
     1741 ctttacggaa gacaatgtat gtatttcggt tcctggagaa actattgcat ctattgcata
     1801 ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt ttccatcttg cacttcaata
     1861 gcatatcttt gttaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     1921 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     1981 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2041 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2101 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2161 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2221 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     2281 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     2341 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     2401 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     2461 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     2521 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     2581 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     2641 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     2701 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     2761 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg
     2821 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcggaata
     2881 ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca
     2941 tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg
     3001 agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt
     3061 aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta
     3121 tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag
     3181 tctcatcctt caatgctatc atttcctttg atattggatc gatccgatga taagctgtca
     3241 aacatgagaa ttgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     3301 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     3361 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     3421 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
     3481 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
     3541 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     3601 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     3661 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     3721 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     3781 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     3841 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     3901 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     3961 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     4021 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     4081 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     4141 tcttcgtttc ctgcatgttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     4201 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     4261 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa
     4321 aaaagaataa aaaaaaaatg atgaattgaa aagctaattc ttgaagacga aagggcctcg
     4381 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4441 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4561 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5101 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5281 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5401 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6301 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6361 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6421 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     6481 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     6541 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     6601 agcttaccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     6661 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     6721 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     6781 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     6841 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     6901 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     6961 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     7021 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
     7081 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
     7141 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
     7201 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
     7261 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
     7321 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
     7381 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
     7441 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
     7501 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
     7561 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
     7621 tcctatcatt atcgtctaac atctaatt
//
