LOCUS       sequence-104905-        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCACS backbone (Amp), SpHIS5, 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 5614)
  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 5614)
  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.HO3.loxP
FEATURES             Location/Qualifiers
     source          1..5614
                     /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"
     gene            2784..3984
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2784..3127
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3128..3781
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      3787..3984
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4023..4056
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4110..4787)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             4794..4805
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4823..4839
                     /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(4847..4853,4854..4871,4872..4877))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4892..4913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    5025..5058
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      join(5594..5614,1..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 tccttgccta acttcaatgt caaagttgaa atattctcct ttagagcgct
     2281 ccatttcttc tatgaagcgt tttgcggcaa actcaccttc aactgtcatt gggaatgtct
     2341 tatgatggtt ttttggaatt attattatcc taccatcaag cgtctgacat tgctgcagat
     2401 ttctccatct cactttatat ttggtggcat ttctaccact tttttccaac agtggtttgg
     2461 tagggaccct gactgacaat ttatgacctg cagtacattg taatgcaaga cgctgataaa
     2521 ctgttctacg cctgggatct aacctaccag gttcaccttc aaaagctctg tgtttggttt
     2581 tttgctgtat attatagatt ttctgatagc cctgtgtgac atttatgacg cgggcagcgg
     2641 agccatctgc gcacataacg taagagttag ccgtgacgtt tgcgatgtct ttaatttcac
     2701 cgttagccat cagaatagtc gtgttttcag aaagcatgga tcctgtttag cttgcctcgt
     2761 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     2821 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     2881 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     2941 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     3001 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     3061 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     3121 aacaaccatg ggtaggaggg cttttgtaga aagaaatacg aacgaaacga aaatcagcgt
     3181 tgccatcgct ttggacaaag ctcccttacc tgaagagtcg aattttattg atgaacttat
     3241 aacttccaag catgcaaacc aaaagggaga acaagtaatc caagtagaca cgggaattgg
     3301 attcttggat cacatgtatc atgcactggc taaacatgca ggctggagct tacgacttta
     3361 ctcaagaggt gatttaatca tcgatgatca tcacactgca gaagatactg ctattgcact
     3421 tggtattgca ttcaagcagg ctatgggtaa ctttgccggc gttaaaagat ttggacatgc
     3481 ttattgtcca cttgacgaag ccctttctag aagcgtagtt gacttgtcgg gacggcccta
     3541 tgctgttatc gatttgggat taaagcgtga aaaggttggg gaattgtcct gtgaaatgat
     3601 ccctcactta ctatattcct tttcggtagc agctggaatt actttgcatg ttacctgctt
     3661 atatggtagt aatgaccatc atcgtgctga aagcgctttt aaatctctgg ctgttgccat
     3721 gcgcgcggct actagtctta ctggaagttc tgaagtccca agcacgaagg gagtgttgta
     3781 aagagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     3841 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     3901 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     3961 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccaggctag
     4021 caataacttc gtataatgta tgctatacga agttatcagt agagaccgcc tggctctagt
     4081 agcgatctac actagcacta tcagcgttat taagcaccgg tggagtgacg accttcagca
     4141 cgttcgtact gttcaacgat ggtgtagtct tcgttgtggg aggtgatgtc cagtttgatg
     4201 tcggttttgt aagcacccgg cagctgaacc ggttttttag ccatgtaggt ggttttaact
     4261 tcagcgtcgt agtgaccacc gtctttcagt ttcagacgca ttttgatttc acctttcaga
     4321 gcaccgtctt ccgggtacat acgttcggtg gaagcttccc aacccatggt ttttttctgc
     4381 ataaccggac cgtcggacgg gaagttggta ccacgcagtt taactttgta gatgaactca
     4441 ccgtcttgca gggaggagtc ctgggtaacg gtaacaacac caccgtcttc gaagttcata
     4501 acacgttccc atttgaaacc ttccgggaag gacagtttca ggtagtccgg gatgtcagcc
     4561 gggtgtttaa cgtaagcttt ggaaccgtac tggaactgcg gggacaggat gtcccaagcg
     4621 aacggcagcg gaccaccttt ggtaactttc agtttagcgg tctgggtacc ttcgtacgga
     4681 cgaccttcac cttcaccttc gatttcgaac tcgtgaccgt taacggaacc ttccatacga
     4741 actttgaaac gcatgaactc tttgataacg tcttcggagg aagccatcta gtatttctcc
     4801 tctttctcta gtatgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4861 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4921 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4981 gccaacgcgc ggggagaggc ggtttgcgta ttgggtctca ttttataact tcgtataatg
     5041 tatgctatac gaagttatac tagtgcatgc tttaaagtat agatagaatt gattgctgct
     5101 tatgaggata tggatttaga gctaatagtt gtaatagaaa taaaagccat ttagaatagg
     5161 aattgaatac aattttatga gctttaaata tttaatgatg aatagtgcgt ttagaacgct
     5221 tcatcatgct tcaacaagct tctgcaaacc tcatttgagt agcaatagct gtttactttt
     5281 ttccgcgact ttattcgaag ttatggtgaa cttttcgtgg tttcacacct aataacgccc
     5341 agcggtagta caattcaaag tagtaggtac caatggtagt actagtgggt gatagtagca
     5401 aatttctcta tgaaaatgaa ttgttgctct gcaatgatgt taatgtggac atgacatgaa
     5461 ttttcgcgat atgttgatgt aaataataac atgattttcg tggatcattt cgtaaaccat
     5521 aggtttattt cgtgaattac taagagaaag gcatagaaac aggacttgcg gatgggatac
     5581 gcggccgctg cagtccggca aaaaagggca aggt
//
