LOCUS       sequence-104859-        5806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS416 backbone, contains an RFP flanked by BsaI sites with CAGT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5806)
  AUTHORS   Agmon N, Mitchell LA, Cai Y, Ikushima S, Chuang J, Zheng A, Choi WJ,
            Martin JA, Caravelli K, Stracquadanio G, Boeke JD
  TITLE     Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae 
            Transcription Units.
  JOURNAL   ACS Synth Biol. 2015 Jul 17;4(7):853-9. doi: 10.1021/sb500372z. Epub
            2015 Mar 23.
  PUBMED    25756291
REFERENCE   2  (bases 1 to 5806)
  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"; doi: "10.1021/sb500372z";
            journalName: "ACS Synth Biol"; date: "2015-07-17- 17"; volume: "4"; 
            issue: "7"; pages: "853-9"
COMMENT     Sequence Label: pAV116
FEATURES             Location/Qualifiers
     source          1..5806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..45
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     145..164
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /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"
     rep_origin      complement(1345..1800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1482..1503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1694..1713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1930..1952
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1944..1961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1945..1961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1971..1990
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1971..1989
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2014..2030
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2015..2031
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2115..2792)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2799..2810
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2828..2844
                     /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)."
     promoter        complement(join(2852..2858,2859..2876,2877..2882))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2897..2918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3033..3049)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3033..3049)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3086..3106)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3086..3104)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3125..3141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3125..3141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3149..3165
                     /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)."
     promoter        complement(join(3173..3179,3180..3197,3198..3203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3356..3373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3527..4115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3607..3626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4286..5077,5078..5146))
                     /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     4909..4928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5147..5251)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5288..5791
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt gtcacagctt
       61 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      181 ccacagcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tgtctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc gcacagatgc
     1321 gtaaggagaa aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt
     1381 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1441 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     1501 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1561 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1621 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1681 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     1741 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1801 cgcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     1861 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     1921 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     1981 cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg
     2041 atatcgaatt cctgcagccc ccagtagaga ccgcctggct ctagtagcga tctacactag
     2101 cactatcagc gttattaagc accggtggag tgacgacctt cagcacgttc gtactgttca
     2161 acgatggtgt agtcttcgtt gtgggaggtg atgtccagtt tgatgtcggt tttgtaagca
     2221 cccggcagct gaaccggttt tttagccatg taggtggttt taacttcagc gtcgtagtga
     2281 ccaccgtctt tcagtttcag acgcattttg atttcacctt tcagagcacc gtcttccggg
     2341 tacatacgtt cggtggaagc ttcccaaccc atggtttttt tctgcataac cggaccgtcg
     2401 gacgggaagt tggtaccacg cagtttaact ttgtagatga actcaccgtc ttgcagggag
     2461 gagtcctggg taacggtaac aacaccaccg tcttcgaagt tcataacacg ttcccatttg
     2521 aaaccttccg ggaaggacag tttcaggtag tccgggatgt cagccgggtg tttaacgtaa
     2581 gctttggaac cgtactggaa ctgcggggac aggatgtccc aagcgaacgg cagcggacca
     2641 cctttggtaa ctttcagttt agcggtctgg gtaccttcgt acggacgacc ttcaccttca
     2701 ccttcgattt cgaactcgtg accgttaacg gaaccttcca tacgaacttt gaaacgcatg
     2761 aactctttga taacgtcttc ggaggaagcc atctagtatt tctcctcttt ctctagtatg
     2821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3001 gaggcggttt gcgtattggg tctcattttg gggatccact agttctagag cggccgccac
     3061 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     3121 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatag
     3181 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa
     3241 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3301 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3361 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3721 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3961 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4201 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4261 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4321 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4381 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac
     4441 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4501 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4561 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4621 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4681 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4741 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4801 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4861 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4921 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4981 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5041 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5101 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5161 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5221 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
     5281 ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa
     5341 attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat
     5401 gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct
     5461 caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct
     5521 gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg
     5581 tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt
     5641 tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac
     5701 aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata
     5761 cgaggcgcgt gtaagttaca ggcaagcgat ccgtcctaag aaacca
//
