LOCUS       sequence-115799-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCACS backbone (Amp), cloNAT, 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 5412)
  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 5412)
  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.KN
FEATURES             Location/Qualifiers
     source          1..5412
                     /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            2722..3841
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        2722..3065
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3066..3638
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      3644..3841
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(3939..4616)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             4623..4634
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4652..4668
                     /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(4676..4682,4683..4700,4701..4706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4721..4742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      join(5392..5412,1..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 cggatcctag gtctagagat ctgtttagct tgcctcgtcc
     2701 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     2761 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     2821 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     2881 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     2941 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     3001 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     3061 caaccatggg taccactctt gacgacacgg cttaccggta ccgcaccagt gtcccggggg
     3121 acgccgaggc catcgaggca ctggatgggt ccttcaccac cgacaccgtc ttccgcgtca
     3181 ccgccaccgg ggacggcttc accctgcggg aggtgccggt ggacccgccc ctgaccaagg
     3241 tgttccccga cgacgaatcg gacgacgaat cggacgacgg ggaggacggc gacccggact
     3301 cccggacgtt cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg gttgtttcgt
     3361 actccggctg gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg gagcaccggg
     3421 ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga gttcgcccgc gagcggggcg
     3481 ccgggcacct ctggctggag gtcaccaacg tcaacgcacc ggcgatccac gcgtaccggc
     3541 ggatggggtt caccctctgc ggcctggaca ccgccctgta cgacggcacc gcctcggacg
     3601 gcgagcaggc gctctacatg agcatgccct gcccctaatc agtactgaca ataaaaagat
     3661 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     3721 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     3781 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     3841 gctgtcgatt cgatactaac gccgccatcc agtgtcgagc tagcacagta gagaccgcct
     3901 ggctctagta gcgatctaca ctagcactat cagcgttatt aagcaccggt ggagtgacga
     3961 ccttcagcac gttcgtactg ttcaacgatg gtgtagtctt cgttgtggga ggtgatgtcc
     4021 agtttgatgt cggttttgta agcacccggc agctgaaccg gttttttagc catgtaggtg
     4081 gttttaactt cagcgtcgta gtgaccaccg tctttcagtt tcagacgcat tttgatttca
     4141 cctttcagag caccgtcttc cgggtacata cgttcggtgg aagcttccca acccatggtt
     4201 tttttctgca taaccggacc gtcggacggg aagttggtac cacgcagttt aactttgtag
     4261 atgaactcac cgtcttgcag ggaggagtcc tgggtaacgg taacaacacc accgtcttcg
     4321 aagttcataa cacgttccca tttgaaacct tccgggaagg acagtttcag gtagtccggg
     4381 atgtcagccg ggtgtttaac gtaagctttg gaaccgtact ggaactgcgg ggacaggatg
     4441 tcccaagcga acggcagcgg accacctttg gtaactttca gtttagcggt ctgggtacct
     4501 tcgtacggac gaccttcacc ttcaccttcg atttcgaact cgtgaccgtt aacggaacct
     4561 tccatacgaa ctttgaaacg catgaactct ttgataacgt cttcggagga agccatctag
     4621 tatttctcct ctttctctag tatgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4681 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4741 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4801 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggtctcat tttactagtg
     4861 catgctttgg acagtgatac cgaagaagga agtaaattca gtagcgccgc atgaggtagt
     4921 agcagaagga gttaaggttg accatggctc acttggcata tactgttggg cagaaacagt
     4981 atagtgaaac tgttatcaaa ccaatatatt tgggctacat ggctttcctt tcccgaaaaa
     5041 gacaccattt agtaattatg ttgtactttt actctctgaa cttctgtgaa tataaaaaat
     5101 atactaaata aaaggatgca acttccttaa agagggcaag atagcgacat aaagtggaaa
     5161 agtgttagca acattgcgca ggcatcgatt aatatatgca tatatacttt ttacgtagtc
     5221 tacgttaaat tgtcaataga tcaagcagaa tcggtagctc tgttaagcga ttaagcccag
     5281 tggctttttg tcgcaccgtt taggttcgcg ttagcaagat ggaaaaaaaa gaacgcgaat
     5341 catgtattga cgtatgacgt attaggcaga tgggatacgc ggccgctgca gtccggcaaa
     5401 aaagggcaag gt
//
