LOCUS       sequence-104877-        5532 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 5532)
  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 5532)
  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.F3.loxP
FEATURES             Location/Qualifiers
     source          1..5532
                     /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            2715..3915
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2715..3058
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3059..3712
                     /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      3718..3915
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3954..3987
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4041..4718)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             4725..4736
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4754..4770
                     /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(4778..4784,4785..4802,4803..4808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    4956..4989
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      join(5512..5532,1..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 tccttgcttg cttctttatt tggcagcatt tttcaaaaat aataaaatgg
     2281 aagccgcgag tacgaacaat gatgtgttct gggaatacct cgtcaaaaca agacaatggc
     2341 aaggattttc tttcatcagg cagaaagatc tggatctgaa tggcatcatt ttgtgatgtg
     2401 taaaagcggg accttgttat ttcgactttt tgcatcatgt tgatgcaatt tgctactttt
     2461 ccgacggtgc gctccaacgg atgggtattt ccttaataac aaggcatttc tctggaagtt
     2521 ggcttactgt ttgaaatcac agccggtcac aaaataaagt aaaaaaacta tctctctcca
     2581 caagaagtaa ttacaggttg tatactacat atgatcgtat ttctttatga acactaagga
     2641 gtttcccgct gtgtaccgca atacccacgg atcctgttta gcttgcctcg tccccgccgg
     2701 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     2761 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     2821 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     2881 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     2941 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     3001 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat
     3061 gggtaggagg gcttttgtag aaagaaatac gaacgaaacg aaaatcagcg ttgccatcgc
     3121 tttggacaaa gctcccttac ctgaagagtc gaattttatt gatgaactta taacttccaa
     3181 gcatgcaaac caaaagggag aacaagtaat ccaagtagac acgggaattg gattcttgga
     3241 tcacatgtat catgcactgg ctaaacatgc aggctggagc ttacgacttt actcaagagg
     3301 tgatttaatc atcgatgatc atcacactgc agaagatact gctattgcac ttggtattgc
     3361 attcaagcag gctatgggta actttgccgg cgttaaaaga tttggacatg cttattgtcc
     3421 acttgacgaa gccctttcta gaagcgtagt tgacttgtcg ggacggccct atgctgttat
     3481 cgatttggga ttaaagcgtg aaaaggttgg ggaattgtcc tgtgaaatga tccctcactt
     3541 actatattcc ttttcggtag cagctggaat tactttgcat gttacctgct tatatggtag
     3601 taatgaccat catcgtgctg aaagcgcttt taaatctctg gctgttgcca tgcgcgcggc
     3661 tactagtctt actggaagtt ctgaagtccc aagcacgaag ggagtgttgt aaagagtact
     3721 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     3781 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     3841 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     3901 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccaggcta gcaataactt
     3961 cgtataatgt atgctatacg aagttatcag tagagaccgc ctggctctag tagcgatcta
     4021 cactagcact atcagcgtta ttaagcaccg gtggagtgac gaccttcagc acgttcgtac
     4081 tgttcaacga tggtgtagtc ttcgttgtgg gaggtgatgt ccagtttgat gtcggttttg
     4141 taagcacccg gcagctgaac cggtttttta gccatgtagg tggttttaac ttcagcgtcg
     4201 tagtgaccac cgtctttcag tttcagacgc attttgattt cacctttcag agcaccgtct
     4261 tccgggtaca tacgttcggt ggaagcttcc caacccatgg tttttttctg cataaccgga
     4321 ccgtcggacg ggaagttggt accacgcagt ttaactttgt agatgaactc accgtcttgc
     4381 agggaggagt cctgggtaac ggtaacaaca ccaccgtctt cgaagttcat aacacgttcc
     4441 catttgaaac cttccgggaa ggacagtttc aggtagtccg ggatgtcagc cgggtgttta
     4501 acgtaagctt tggaaccgta ctggaactgc ggggacagga tgtcccaagc gaacggcagc
     4561 ggaccacctt tggtaacttt cagtttagcg gtctgggtac cttcgtacgg acgaccttca
     4621 ccttcacctt cgatttcgaa ctcgtgaccg ttaacggaac cttccatacg aactttgaaa
     4681 cgcatgaact ctttgataac gtcttcggag gaagccatct agtatttctc ctctttctct
     4741 agtatgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4801 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4861 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4921 cggggagagg cggtttgcgt attgggtctc attttataac ttcgtataat gtatgctata
     4981 cgaagttata ctagtgcatg cagaaacttc tgtggcttga cagataaata actgcagtag
     5041 tcggtgcgta ctaattgttt ggtcgtgtct gaaaaatctt gaattttcag aaaagaataa
     5101 gccccaaatg tcagtgatgg tagtagcagt actcccctac gattttagat actttagaga
     5161 gcccaccttc agaatcggaa ggaggataat tttgtaaagc ccttctgttt tttctcttgc
     5221 ataacttata tttccacatc aaaaagtagt gtgctaagaa aaaggtcaca agaaaaagga
     5281 ttacggcact ctctgcatct agacatatac caaaagttgg gtttgctcac gaaaatacca
     5341 taattgtggt gtcaaaaaaa tcctgcctca taataccact gcagcaattg tggatgacta
     5401 aaaaataact tgcattccac gatgttattt tactttataa agcacctgca attttttttt
     5461 ttgtattaac tcatcgagta tgtctgatga tgggatacgc ggccgctgca gtccggcaaa
     5521 aaagggcaag gt
//
