LOCUS       sequence-121508-        5171 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 5171)
  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 5171)
  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.loxP
FEATURES             Location/Qualifiers
     source          1..5171
                     /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"
     protein_bind    1319..1352
                     /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(1432..2109)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2116..2127
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2145..2161
                     /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(2169..2175,2176..2193,2194..2199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2214..2235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    2365..2398
                     /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(2451..2471)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2451..2469)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2490..2506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2490..2506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2514..2530
                     /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(2538..2544,2545..2562,2563..2568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2721..2738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2892..3480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2972..2991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3651..4442,4443..4511))
                     /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     4274..4293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4512..4616)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4653..5156
                     /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 ccctcctcga gcgtcgacgg cagctgggat
     1321 aacttcgtat aatgtacatt atacgaagtt atgggacgtg gcgaattcct gcagccccca
     1381 gtagagaccg cctggctcta gtagcgatct acactagcac tatcagcgtt attaagcacc
     1441 ggtggagtga cgaccttcag cacgttcgta ctgttcaacg atggtgtagt cttcgttgtg
     1501 ggaggtgatg tccagtttga tgtcggtttt gtaagcaccc ggcagctgaa ccggtttttt
     1561 agccatgtag gtggttttaa cttcagcgtc gtagtgacca ccgtctttca gtttcagacg
     1621 cattttgatt tcacctttca gagcaccgtc ttccgggtac atacgttcgg tggaagcttc
     1681 ccaacccatg gtttttttct gcataaccgg accgtcggac gggaagttgg taccacgcag
     1741 tttaactttg tagatgaact caccgtcttg cagggaggag tcctgggtaa cggtaacaac
     1801 accaccgtct tcgaagttca taacacgttc ccatttgaaa ccttccggga aggacagttt
     1861 caggtagtcc gggatgtcag ccgggtgttt aacgtaagct ttggaaccgt actggaactg
     1921 cggggacagg atgtcccaag cgaacggcag cggaccacct ttggtaactt tcagtttagc
     1981 ggtctgggta ccttcgtacg gacgaccttc accttcacct tcgatttcga actcgtgacc
     2041 gttaacggaa ccttccatac gaactttgaa acgcatgaac tctttgataa cgtcttcgga
     2101 ggaagccatc tagtatttct cctctttctc tagtatgtgt gaaattgtta tccgctcaca
     2161 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2221 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2281 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggtct
     2341 cattttgggg atcccggagc acccataact tcgtataatg tacattatac gaagttatcc
     2401 caggtcgctc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     2461 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2521 ccgctcacaa ttccacacaa cataggagcc ggaagcataa agtgtaaagc ctggggtgcc
     2581 taatgagtga ggtaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2641 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2701 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2761 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3601 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3661 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3721 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3781 atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3841 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3901 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3961 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4021 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4081 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4141 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4201 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4261 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4321 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4381 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4441 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4501 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4561 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4621 tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat aattataatt
     4681 taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga aattaaagaa
     4741 aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca acaaatacta
     4801 ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca tcttgtctgt
     4861 gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt aatcgaatgt
     4921 atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt
     4981 gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt
     5041 tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc
     5101 caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc
     5161 ctaagaaacc a
//
