LOCUS       sequence-104885-        5696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS414 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 5696)
  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 5696)
  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: pAV114
FEATURES             Location/Qualifiers
     source          1..5696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..22
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     122..141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        158..438
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             439..1113
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1212..1667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1349..1370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1561..1580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1797..1819
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1811..1828
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1812..1828
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1838..1857
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1838..1856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1881..1897
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1882..1898
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1982..2659)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2666..2677
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2695..2711
                     /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(2719..2725,2726..2743,2744..2749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2764..2785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2900..2916)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2900..2916)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2953..2973)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2953..2971)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2992..3008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2992..3008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3016..3032
                     /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(3040..3046,3047..3064,3065..3070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3223..3240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3394..3982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3474..3493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4153..4944,4945..5013))
                     /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     4776..4795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5014..5118)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5155..5658
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaaaataggc gtatcacgag gccctttgtc acagcttgtc tgtaagcgga tgccgggagc
       61 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
      121 gcggcatcag agcagattgt actgagagtg caccataaac gacattacta tatatataat
      181 ataggaagca tttaatagac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag
      241 atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt
      301 gatccggagc ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga
      361 gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac
      421 acaaaggcag cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg
      481 aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac
      541 ttgctgggta ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg
      601 aaaatttcaa gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt
      661 ggcgtgtttc gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt
      721 gatatcgtcc aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg
      781 ccagttatta aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca
      841 cagaaacctc attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt
      901 ttggattgga actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat
      961 tttatgttag ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc
     1021 gttattggtg ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa
     1081 atagcaaatt tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag
     1141 tattgtttgt gcacttgcct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1201 accgcatcag gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1261 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1321 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1381 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1441 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1501 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1561 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1621 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc gccattcgcc
     1681 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     1741 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     1801 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     1861 attgggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct
     1921 gcagccccca gtagagaccg cctggctcta gtagcgatct acactagcac tatcagcgtt
     1981 attaagcacc ggtggagtga cgaccttcag cacgttcgta ctgttcaacg atggtgtagt
     2041 cttcgttgtg ggaggtgatg tccagtttga tgtcggtttt gtaagcaccc ggcagctgaa
     2101 ccggtttttt agccatgtag gtggttttaa cttcagcgtc gtagtgacca ccgtctttca
     2161 gtttcagacg cattttgatt tcacctttca gagcaccgtc ttccgggtac atacgttcgg
     2221 tggaagcttc ccaacccatg gtttttttct gcataaccgg accgtcggac gggaagttgg
     2281 taccacgcag tttaactttg tagatgaact caccgtcttg cagggaggag tcctgggtaa
     2341 cggtaacaac accaccgtct tcgaagttca taacacgttc ccatttgaaa ccttccggga
     2401 aggacagttt caggtagtcc gggatgtcag ccgggtgttt aacgtaagct ttggaaccgt
     2461 actggaactg cggggacagg atgtcccaag cgaacggcag cggaccacct ttggtaactt
     2521 tcagtttagc ggtctgggta ccttcgtacg gacgaccttc accttcacct tcgatttcga
     2581 actcgtgacc gttaacggaa ccttccatac gaactttgaa acgcatgaac tctttgataa
     2641 cgtcttcgga ggaagccatc tagtatttct cctctttctc tagtatgtgt gaaattgtta
     2701 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2761 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2821 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2881 tattgggtct cattttgggg atccactagt tctagagcgg ccgccaccgc ggtggagctc
     2941 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     3001 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
     3061 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc
     3121 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4081 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4141 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4201 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4261 accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt
     4321 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4381 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4441 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4501 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4561 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4621 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4681 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4741 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4801 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4861 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4921 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4981 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5041 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5101 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg
     5161 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
     5221 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
     5281 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
     5341 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
     5401 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
     5461 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
     5521 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
     5581 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
     5641 agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta acctat
//
