LOCUS       sequence-115802-        5694 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 5694)
  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 5694)
  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.HO6
FEATURES             Location/Qualifiers
     source          1..5694
                     /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"
     CDS             complement(4224..4901)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             4908..4919
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4937..4953
                     /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(4961..4967,4968..4985,4986..4991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5006..5027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      join(5674..5694,1..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 tccagctcgt tttatttagg
     2761 ttctatcgag gagaaaaagc gacaagaaga gatagaccat ggataaactg attatgttct
     2821 aaacactcct cagaagctca tcgaactgtc atcctgcgtg aagattaaaa tccaacttag
     2881 aaatttcgag cttacggaga caatcatatg ggagaagcaa ttggaagata gaaaaaaggt
     2941 actcggtaca taaatatatg tgattctggg tagaagatcg gtctgcattg gatggtggta
     3001 acgcattttt ttacacacat tacttgcctc gagcatcaaa tggtggttat tcgtggatct
     3061 atatcacgtg atttgcttaa gaattgtcgt tcatggtgac acttttagct ttgacatgat
     3121 taagctcatc tcaattgatg ttatctaaag tcatttcaac tatctaagat gtggttgtga
     3181 ttgggccatt ttgtgaaagc cagtacgcca gcgtcaatac actcccgtca attagttgca
     3241 ccatgtccac aaaatcatat accagtagag ctgagactca tgcaagtccg gttgcatcga
     3301 aacttttacg tttaatggat gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt
     3361 cgactgatga gctattgaaa cttgttgaaa cgttgggtcc atacatttgc cttttgaaaa
     3421 cacacgttga tatcttggat gatttcagtt atgagggtac tgtcgttcca ttgaaagcat
     3481 tggcagagaa atacaagttc ttgatatttg aggacagaaa attcgccgat atcggtaaca
     3541 cagtcaaatt acaatataca tcgggcgttt accgtatcgc agaatggtct gatatcacca
     3601 acgcccacgg ggttactggt gctggtattg ttgctggctt gaaacaaggt gcgcaagagg
     3661 tcaccaaaga accaagggga ttattgatgc ttgctgaatt gtcttccaag ggttctctag
     3721 cacacggtga atatactaag ggtaccgttg atattgcaaa gagtgataaa gatttcgtta
     3781 ttgggttcat tgctcagaac gatatgggag gaagagaaga agggtttgat tggctaatca
     3841 tgaccccagg tgtaggttta gacgacaaag gcgatgcatt gggtcagcag tacagaaccg
     3901 tcgacgaagt tgtaagtggt ggatcagata tcatcattgt tggcagagga cttttcgcca
     3961 agggtagaga tcctaaggtt gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc
     4021 aaaagagaat cagcgctccc cattaattat acaggaaact taatagaaca aatcacatat
     4081 ttaatctaat agccacctgc attggcacgg tgcaacacta cttcaacttc atcttacaaa
     4141 aagatcacgt gatctgttgt attgtgctag cagtagagac cgcctggctc tagtagcgat
     4201 ctacactagc actatcagcg ttattaagca ccggtggagt gacgaccttc agcacgttcg
     4261 tactgttcaa cgatggtgta gtcttcgttg tgggaggtga tgtccagttt gatgtcggtt
     4321 ttgtaagcac ccggcagctg aaccggtttt ttagccatgt aggtggtttt aacttcagcg
     4381 tcgtagtgac caccgtcttt cagtttcaga cgcattttga tttcaccttt cagagcaccg
     4441 tcttccgggt acatacgttc ggtggaagct tcccaaccca tggttttttt ctgcataacc
     4501 ggaccgtcgg acgggaagtt ggtaccacgc agtttaactt tgtagatgaa ctcaccgtct
     4561 tgcagggagg agtcctgggt aacggtaaca acaccaccgt cttcgaagtt cataacacgt
     4621 tcccatttga aaccttccgg gaaggacagt ttcaggtagt ccgggatgtc agccgggtgt
     4681 ttaacgtaag ctttggaacc gtactggaac tgcggggaca ggatgtccca agcgaacggc
     4741 agcggaccac ctttggtaac tttcagttta gcggtctggg taccttcgta cggacgacct
     4801 tcaccttcac cttcgatttc gaactcgtga ccgttaacgg aaccttccat acgaactttg
     4861 aaacgcatga actctttgat aacgtcttcg gaggaagcca tctagtattt ctcctctttc
     4921 tctagtatgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5101 gcgcggggag aggcggtttg cgtattgggt ctcattttac tagtgcatgc tttaaagtat
     5161 agatagaatt gattgctgct tatgaggata tggatttaga gctaatagtt gtaatagaaa
     5221 taaaagccat ttagaatagg aattgaatac aattttatga gctttaaata tttaatgatg
     5281 aatagtgcgt ttagaacgct tcatcatgct tcaacaagct tctgcaaacc tcatttgagt
     5341 agcaatagct gtttactttt ttccgcgact ttattcgaag ttatggtgaa cttttcgtgg
     5401 tttcacacct aataacgccc agcggtagta caattcaaag tagtaggtac caatggtagt
     5461 actagtgggt gatagtagca aatttctcta tgaaaatgaa ttgttgctct gcaatgatgt
     5521 taatgtggac atgacatgaa ttttcgcgat atgttgatgt aaataataac atgattttcg
     5581 tggatcattt cgtaaaccat aggtttattt cgtgaattac taagagaaag gcatagaaac
     5641 aggacttgcg gatgggatac gcggccgctg cagtccggca aaaaagggca aggt
//
