LOCUS       sequence-104913-        5914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCACS backbone (Amp), TRP1, 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 5914)
  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 5914)
  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: pAV10.K4
FEATURES             Location/Qualifiers
     source          1..5914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        1214..1494
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1495..2169
                     /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"
     CDS             complement(2231..2908)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2915..2926
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2944..2960
                     /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(2968..2974,2975..2992,2993..2998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3013..3034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3683..3699)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3684..3700)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3729..3747)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3768..3784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3768..3784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3792..3808
                     /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(3816..3822,3823..3840,3841..3846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3861..3882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3999..4016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4170..4758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4250..4269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4929..5720,5721..5789))
                     /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     5552..5571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5790..5894)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atccccctta attaaagtct cttgtttacg
      721 ttacactggc aattttaacg gcgagactca tctgaaaaaa gctattcgac ttagtcctgt
      781 gtttttcatt acagataatc ctgatcaacg ggtgtacgtt tttccatttt aggtcagtat
      841 cctccctctg tgtttcaaac taagtataag cgcccaccct aattcgctcg tatccttgtg
      901 tcaacttctt tgctgctctg cagctgtaaa cagtttttgc gataagcctc acatatgtca
      961 gtgctgtaaa taccggaaat agaaagtcac tgtggtctgt accctcattt gatttacgcg
     1021 gttagaaaag tggaaactct ttcttaaccg tgtaaatcaa gtatgtcatt tcaggaaact
     1081 tcctcaattc ttggcgtact atattgagtt gccaatgtat gagagtggtc gatttcaatc
     1141 ggcaacattc tatctcgcaa ttacgcaata aaaattgaca tggtatgtta ttttcatcct
     1201 tcggaggccg gccaacgaca ttactatata tataatatag gaagcattta atagacagca
     1261 tcgtaatata tgtgtacttt gcagttatga cgccagatgg cagtagtgga agatattctt
     1321 tattgaaaaa tagcttgtca ccttacgtac aatcttgatc cggagctttt ctttttttgc
     1381 cgattaagaa ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg agggcattgg
     1441 tgactattga gcacgtgagt atacgtgatt aagcacacaa aggcagcttg gagtatgtct
     1501 gttattaatt tcacaggtag ttctggtcca ttggtgaaag tttgcggctt gcagagcaca
     1561 gaggccgcag aatgtgctct agattccgat gctgacttgc tgggtattat atgtgtgccc
     1621 aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca
     1681 tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag
     1741 gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat
     1801 gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt
     1861 ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc
     1921 ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac
     1981 tgggttggaa ggcaagagag ccccgaaagc ttacatttta tgttagctgg tggactgacg
     2041 ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga
     2101 ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct
     2161 aagaaatagg gccggcccag tagagaccgc ctggctctag tagcgatcta cactagcact
     2221 atcagcgtta ttaagcaccg gtggagtgac gaccttcagc acgttcgtac tgttcaacga
     2281 tggtgtagtc ttcgttgtgg gaggtgatgt ccagtttgat gtcggttttg taagcacccg
     2341 gcagctgaac cggtttttta gccatgtagg tggttttaac ttcagcgtcg tagtgaccac
     2401 cgtctttcag tttcagacgc attttgattt cacctttcag agcaccgtct tccgggtaca
     2461 tacgttcggt ggaagcttcc caacccatgg tttttttctg cataaccgga ccgtcggacg
     2521 ggaagttggt accacgcagt ttaactttgt agatgaactc accgtcttgc agggaggagt
     2581 cctgggtaac ggtaacaaca ccaccgtctt cgaagttcat aacacgttcc catttgaaac
     2641 cttccgggaa ggacagtttc aggtagtccg ggatgtcagc cgggtgttta acgtaagctt
     2701 tggaaccgta ctggaactgc ggggacagga tgtcccaagc gaacggcagc ggaccacctt
     2761 tggtaacttt cagtttagcg gtctgggtac cttcgtacgg acgaccttca ccttcacctt
     2821 cgatttcgaa ctcgtgaccg ttaacggaac cttccatacg aactttgaaa cgcatgaact
     2881 ctttgataac gtcttcggag gaagccatct agtatttctc ctctttctct agtatgtgtg
     2941 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3001 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3121 cggtttgcgt attgggtctc attttttcaa accattctgt tataagagat atgtaccatc
     3181 aatcgccgac agcgataccc tctttaacct ctagtaagaa ggacccggca cttattgtaa
     3241 tacgtttagg ggaagtaggt aaatctggac cggtaatacg gcataacgat ccatttcgga
     3301 catcggggac atttcttttt tttcctttat tatttttttt ttctaatgtc cttacgaatt
     3361 tcaggctcta accttatttc ttattaggac gtgtactcat tgccatcgca acaataaaaa
     3421 taaaacatta ataaaaaatt aattagagcc tgtaacatag agatcaacgc tactgtagcg
     3481 ttagaaggac agtgagagag tggcatgaca attaaagttt cttccgatta cttcatgtca
     3541 tctgcattac aaaggaagga aagatgtttc gaattatttc ccctttaagg tatttttgat
     3601 gtgaaatttg accccatact gtgaaaatca actgctaaga actctttaat taagggctgc
     3661 aggaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc ggtacccagc
     3721 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
     3781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3841 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3901 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3961 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4021 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4801 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4861 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4921 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4981 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5041 tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca
     5101 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5161 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5221 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5281 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5341 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5401 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5461 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5521 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5581 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5641 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5701 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5761 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5821 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5881 ataggggttc cgcgcacatt tccccgaaaa gtgc
//
