LOCUS       sequence-104900-        5690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCACS backbone (Amp), KlURA3, 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 5690)
  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 5690)
  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.K6.loxP
FEATURES             Location/Qualifiers
     source          1..5690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..78)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(232..820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(312..331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1003..1097)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             join(1115..1183,1184..1975)
                     /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     complement(1333..1352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2146..2164
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2168..2211)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    4096..4129
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4183..4860)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             4867..4878
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4896..4912
                     /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(4920..4926,4927..4944,4945..4950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4965..4986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    5098..5131
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      join(5670..5690,1..37)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
ORIGIN
        1 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc
       61 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag
      241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      961 aagtatatat gagtaaactt ggtctgacag ctcgaggctt ggattctcac caataaaaaa
     1021 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     1081 atctatcaac aggagtccaa gcgagctcga tatcatgagt attcaacatt tccgtgtcgc
     1141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1621 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1801 tgctgataaa tctggagccg gtgagcgtgg atctcgcggt atcattgcag cactggggcc
     1861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1921 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaatttct
     1981 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     2041 atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaac tcgagtgcca
     2101 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2161 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
     2221 cggccgcgta tccttgcgtt ctctaatcct ttcgtcgtgt atattgtaaa aactgctgaa
     2281 cgagtattga taaaagatat cttggccact aaggggcaga tccccttctg gtgtgataga
     2341 caaccccagg agcatagata acaccaactt gtggtggagg gtcatcgaat tggaattgtc
     2401 tgttggcaac agtagaacag atgctgccct tgctatctgt caaaatgccg ctcttcaaag
     2461 ttactttcaa agcgctgtca ctgctacgac agcttttttt ttagaaacag cagcaatgcc
     2521 ttgacatgta acgtaagaaa agaaaaaaga gatggcagaa gaaatactaa gcgataacgg
     2581 caatgtagag gtgctttttt tatcggaata aatagagaag tcagtaacag tgattgctgt
     2641 ggctccctct ttaatcgtat cggatccagc tcgttttatt taggttctat cgaggagaaa
     2701 aagcgacaag aagagataga ccatggataa actgattatg ttctaaacac tcctcagaag
     2761 ctcatcgaac tgtcatcctg cgtgaagatt aaaatccaac ttagaaattt cgagcttacg
     2821 gagacaatca tatgggagaa gcaattggaa gatagaaaaa aggtactcgg tacataaata
     2881 tatgtgattc tgggtagaag atcggtctgc attggatggt ggtaacgcat ttttttacac
     2941 acattacttg cctcgagcat caaatggtgg ttattcgtgg atctatatca cgtgatttgc
     3001 ttaagaattg tcgttcatgg tgacactttt agctttgaca tgattaagct catctcaatt
     3061 gatgttatct aaagtcattt caactatcta agatgtggtt gtgattgggc cattttgtga
     3121 aagccagtac gccagcgtca atacactccc gtcaattagt tgcaccatgt ccacaaaatc
     3181 atataccagt agagctgaga ctcatgcaag tccggttgca tcgaaacttt tacgtttaat
     3241 ggatgaaaag aagaccaatt tgtgtgcttc tcttgacgtt cgttcgactg atgagctatt
     3301 gaaacttgtt gaaacgttgg gtccatacat ttgccttttg aaaacacacg ttgatatctt
     3361 ggatgatttc agttatgagg gtactgtcgt tccattgaaa gcattggcag agaaatacaa
     3421 gttcttgata tttgaggaca gaaaattcgc cgatatcggt aacacagtca aattacaata
     3481 tacatcgggc gtttaccgta tcgcagaatg gtctgatatc accaacgccc acggggttac
     3541 tggtgctggt attgttgctg gcttgaaaca aggtgcgcaa gaggtcacca aagaaccaag
     3601 gggattattg atgcttgctg aattgtcttc caagggttct ctagcacacg gtgaatatac
     3661 taagggtacc gttgatattg caaagagtga taaagatttc gttattgggt tcattgctca
     3721 gaacgatatg ggaggaagag aagaagggtt tgattggcta atcatgaccc caggtgtagg
     3781 tttagacgac aaaggcgatg cattgggtca gcagtacaga accgtcgacg aagttgtaag
     3841 tggtggatca gatatcatca ttgttggcag aggacttttc gccaagggta gagatcctaa
     3901 ggttgaaggt gaaagataca gaaatgctgg atgggaagcg taccaaaaga gaatcagcgc
     3961 tccccattaa ttatacagga aacttaatag aacaaatcac atatttaatc taatagccac
     4021 ctgcattggc acggtgcaac actcacttca acttcatctt acaaaagatc acgtgatctg
     4081 ttgtattggc tagcaataac ttcgtataat gtatgctata cgaagttatc agtagagacc
     4141 gcctggctct agtagcgatc tacactagca ctatcagcgt tattaagcac cggtggagtg
     4201 acgaccttca gcacgttcgt actgttcaac gatggtgtag tcttcgttgt gggaggtgat
     4261 gtccagtttg atgtcggttt tgtaagcacc cggcagctga accggttttt tagccatgta
     4321 ggtggtttta acttcagcgt cgtagtgacc accgtctttc agtttcagac gcattttgat
     4381 ttcacctttc agagcaccgt cttccgggta catacgttcg gtggaagctt cccaacccat
     4441 ggtttttttc tgcataaccg gaccgtcgga cgggaagttg gtaccacgca gtttaacttt
     4501 gtagatgaac tcaccgtctt gcagggagga gtcctgggta acggtaacaa caccaccgtc
     4561 ttcgaagttc ataacacgtt cccatttgaa accttccggg aaggacagtt tcaggtagtc
     4621 cgggatgtca gccgggtgtt taacgtaagc tttggaaccg tactggaact gcggggacag
     4681 gatgtcccaa gcgaacggca gcggaccacc tttggtaact ttcagtttag cggtctgggt
     4741 accttcgtac ggacgacctt caccttcacc ttcgatttcg aactcgtgac cgttaacgga
     4801 accttccata cgaactttga aacgcatgaa ctctttgata acgtcttcgg aggaagccat
     4861 ctagtatttc tcctctttct ctagtatgtg tgaaattgtt atccgctcac aattccacac
     4921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5041 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggtc tcattttata
     5101 acttcgtata atgtatgcta tacgaagtta tactagtgca tgctttggac agtgataccg
     5161 aagaaggaag taaattcagt agcgccgcat gaggtagtag cagaaggagt taaggttgac
     5221 catggctcac ttggcatata ctgttgggca gaaacagtat agtgaaactg ttatcaaacc
     5281 aatatatttg ggctacatgg ctttcctttc ccgaaaaaga caccatttag taattatgtt
     5341 gtacttttac tctctgaact tctgtgaata taaaaaatat actaaataaa aggatgcaac
     5401 ttccttaaag agggcaagat agcgacataa agtggaaaag tgttagcaac attgcgcagg
     5461 catcgattaa tatatgcata tatacttttt acgtagtcta cgttaaattg tcaatagatc
     5521 aagcagaatc ggtagctctg ttaagcgatt aagcccagtg gctttttgtc gcaccgttta
     5581 ggttcgcgtt agcaagatgg aaaaaaaaga acgcgaatca tgtattgacg tatgacgtat
     5641 taggcagatg ggatacgcgg ccgctgcagt ccggcaaaaa agggcaaggt
//
