LOCUS       sequence-131462-        5963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS413 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 5963)
  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 5963)
  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: pAV113.loxP
FEATURES             Location/Qualifiers
     source          1..5963
                     /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        311..498
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             499..1161
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1421..1876)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1558..1579)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1770..1789
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2006..2028
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2020..2037
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2021..2037
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2047..2066
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2047..2065
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2111..2144
                     /label=loxPsym
                     /bound_moiety="Cre recombinase"
                     /note="loxPsym has a symmetric spacer region, and loxPsym 
                     sites are capable of recombining in either orientation 
                     (Hoess et al., 1986)."
     CDS             complement(2224..2901)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2908..2919
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2937..2953
                     /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(2961..2967,2968..2985,2986..2991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3006..3027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    3157..3190
                     /label=loxPsym
                     /bound_moiety="Cre recombinase"
                     /note="loxPsym has a symmetric spacer region, and loxPsym 
                     sites are capable of recombining in either orientation 
                     (Hoess et al., 1986)."
     primer_bind     complement(3243..3263)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3243..3261)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3282..3298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3282..3298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3306..3322
                     /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(3330..3336,3337..3354,3355..3360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3375..3396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3513..3530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3684..4272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3764..3783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4443..5234,5235..5303))
                     /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     5066..5085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5304..5408)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5445..5948
                     /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 cgtgttggcg
      121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      181 aattcccgtt ttaagagctt ggtgagcgct aggagtcact gccaggtatc gtttgaacac
      241 ggcattagtc agggaagtca taacacagtc ctttcccgca attttctttt tctattactc
      301 ttggcctcct ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt
      361 ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa
      421 ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca
      481 agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa
      541 accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc
      601 ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac
      661 gtccacacag gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc
      721 tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgaa
      781 gactgcggga ttgctctcgg tcaagctttt aaagaggccc taggggccgt gcgtggagta
      841 aaaaggtttg gatcaggatt tgcgcctttg gatgaggcac tttccagagc ggtggtagat
      901 ctttcgaaca ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa agtaggagat
      961 ctctcttgcg agatgatccc gcattttctt gaaagctttg cagaggctag cagaattacc
     1021 ctccacgttg attgtctgcg aggcaagaat gatcatcacc gtagtgagag tgcgttcaag
     1081 gctcttgcgg ttgccataag agaagccacc tcgcccaatg gtaccaacga tgttccctcc
     1141 accaaaggtg ttcttatgta gtgacaccga ttatttaaag ctgcagcata cgatatatat
     1201 acatgtgtat atatgtatac ctatgaatgt cagtaagtat gtatacgaac agtatgatac
     1261 tgaagatgac aaggtaatgc atcattctat acgtgtcatt ctgaacgagg cgcgctttcc
     1321 ttttttcttt ttgctttttc tttttttttc tcttgaactc gacggatcta tgcggtgtga
     1381 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1441 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1501 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1561 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1621 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1681 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1741 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1801 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1861 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1921 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1981 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     2041 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac
     2101 ggcagctggg ataacttcgt ataatgtaca ttatacgaag ttatgggacg tggcgaattc
     2161 ctgcagcccc cagtagagac cgcctggctc tagtagcgat ctacactagc actatcagcg
     2221 ttattaagca ccggtggagt gacgaccttc agcacgttcg tactgttcaa cgatggtgta
     2281 gtcttcgttg tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac ccggcagctg
     2341 aaccggtttt ttagccatgt aggtggtttt aacttcagcg tcgtagtgac caccgtcttt
     2401 cagtttcaga cgcattttga tttcaccttt cagagcaccg tcttccgggt acatacgttc
     2461 ggtggaagct tcccaaccca tggttttttt ctgcataacc ggaccgtcgg acgggaagtt
     2521 ggtaccacgc agtttaactt tgtagatgaa ctcaccgtct tgcagggagg agtcctgggt
     2581 aacggtaaca acaccaccgt cttcgaagtt cataacacgt tcccatttga aaccttccgg
     2641 gaaggacagt ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag ctttggaacc
     2701 gtactggaac tgcggggaca ggatgtccca agcgaacggc agcggaccac ctttggtaac
     2761 tttcagttta gcggtctggg taccttcgta cggacgacct tcaccttcac cttcgatttc
     2821 gaactcgtga ccgttaacgg aaccttccat acgaactttg aaacgcatga actctttgat
     2881 aacgtcttcg gaggaagcca tctagtattt ctcctctttc tctagtatgt gtgaaattgt
     2941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3121 cgtattgggt ctcattttgg ggatcccgga gcacccataa cttcgtataa tgtacattat
     3181 acgaagttat cccaggtcgc tctagagcgg ccgccaccgc ggtggagctc cagcttttgt
     3241 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     3301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4081 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4561 cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata
     4621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5401 gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa
     5461 ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa
     5521 gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc
     5581 caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt
     5641 catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg
     5701 ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt
     5761 acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc
     5821 ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac
     5881 agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc
     5941 aagcgatccg tcctaagaaa cca
//
