LOCUS       sequence-267056-        7018 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 7018)
  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 7018)
  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.F6.loxP
FEATURES             Location/Qualifiers
     source          1..7018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    146..167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(182..187,188..205,206..212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    220..236
                     /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)."
     RBS             254..265
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             272..949
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     protein_bind    complement(1003..1036)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      2927..2970
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2974..2992)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(3163..3954,3955..4023))
                     /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     3786..3805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5656..6244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6145..6164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6398..6415
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6439..6496)
                     /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 ataacttcgt atagcataca ttatacgaag ttataaaatg agacccaata cgcaaaccgc
       61 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      121 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      181 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      241 acacatacta gagaaagagg agaaatacta gatggcttcc tccgaagacg ttatcaaaga
      301 gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga
      361 aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa
      421 aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa
      481 agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg
      541 tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga
      601 ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc
      661 gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat
      721 gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg
      781 tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc
      841 gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat
      901 cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat aacgctgata
      961 gtgctagtgt agatcgctac tagagccagg cggtctctac tgataacttc gtatagcata
     1021 cattatacga agttattgct agccaataca acagatcacg tgatcttttt gtaagatgaa
     1081 gttgaagtag tgttgcaccg tgccaatgca ggtggctatt agattaaata tgtgatttgt
     1141 tctattaagt ttcctgtata attaatgggg agcgctgatt ctcttttggt acgcttccca
     1201 tccagcattt ctgtatcttt caccttcaac cttaggatct ctacccttgg cgaaaagtcc
     1261 tctgccaaca atgatgatat ctgatccacc acttacaact tcgtcgacgg ttctgtactg
     1321 ctgacccaat gcatcgcctt tgtcgtctaa acctacacct ggggtcatga ttagccaatc
     1381 aaacccttct tctcttcctc ccatatcgtt ctgagcaatg aacccaataa cgaaatcttt
     1441 atcactcttt gcaatatcaa cggtaccctt agtatattca ccgtgtgcta gagaaccctt
     1501 ggaagacaat tcagcaagca tcaataatcc ccttggttct ttggtgacct cttgcgcacc
     1561 ttgtttcaag ccagcaacaa taccagcacc agtaaccccg tgggcgttgg tgatatcaga
     1621 ccattctgcg atacggtaaa cgcccgatgt atattgtaat ttgactgtgt taccgatatc
     1681 ggcgaatttt ctgtcctcaa atatcaagaa cttgtatttc tctgccaatg ctttcaatgg
     1741 aacgacagta ccctcataac tgaaatcatc caagatatca acgtgtgttt tcaaaaggca
     1801 aatgtatgga cccaacgttt caacaagttt caatagctca tcagtcgaac gaacgtcaag
     1861 agaagcacac aaattggtct tcttttcatc cattaaacgt aaaagtttcg atgcaaccgg
     1921 actcgcatga gtctcagctc tactggtata tgattttgtg gacatggtgc aactaattga
     1981 cgggagtgta ttgacgctgg cgtactggct ttcacaaaat ggcccaatca caaccacatc
     2041 ttagatagtt gaaatgactt tagataacat caattgagat gagcttaatc atgtcaaagc
     2101 taaaagtgtc accatgaacg acaattctta agcaaatcac gtgatataga tccacgaata
     2161 accaccattt gatgctcgag gcaagtaatg tgtgtaaaaa aatgcgttac caccatccaa
     2221 tgcagaccga tcttctaccc agaatcacat atatttatgt accgagtacc ttttttctat
     2281 cttccaattg cttctcccat atgattgtct ccgtaagctc gaaatttcta agttggattt
     2341 taatcttcac gcaggatgac agttcgatga gcttctgagg agtgtttaga acataatcat
     2401 ttatccatgg tctatctctt cttgtcgctt tttctcctcg atagaaccta aataaaacga
     2461 gctggatccg tggatattgc ggtacacagc gggaaactcc ttagtgttca taaagaaata
     2521 cgatcatatg tagtatacaa cctgtaatta cttcttgtgg agagagatag tttttttact
     2581 ttattttgtg accggctgtg atttcaaaca gtaagccaac ttccagagaa atgccttgtt
     2641 attaaggaaa tacccatccg ttggagcgca ccgtcggaaa agtagcaaat tgcatcaaca
     2701 tgatgcaaaa agtcgaaata acaaggtccc gcttttacac atcacaaaat gatgccattc
     2761 agatccagat ctttctgcct gatgaaagaa aatccttgcc attgtcttgt tttgacgagg
     2821 tattcccaga acacatcatt gttcgtactc gcggcttcca ttttattatt tttgaaaaat
     2881 gctgccaaat aaagaagcaa gcaaggatac gcggccgcga attccagaaa tcatccttag
     2941 cgaaagctaa ggattttttt tatctgaaat tctgcctcgt gatacgccta tttttatagg
     3001 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cactcgagtt gatcgggcac
     3061 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     3121 ttatcgagat tttcaggagc taaggaagct aaaatggaga aattaccaat gcttaatcag
     3181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3241 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3301 gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3361 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3421 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3481 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3541 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3601 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3661 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3721 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3781 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3841 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3901 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3961 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4021 catgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     4081 tctggatttg ttcagaacgc tcggttgccg ccgggcgctg atgaatcccc taatgatttt
     4141 ggtaaaaatc attaagttaa ggtggataca catcttgtca tatgatcaaa tggtttcgcg
     4201 aaaaatcaat aatcagacaa caagatgtgc gaactcgata ttttacacga ctctctttac
     4261 caattctgcc ccgaattaca cttaaaacga ctcaacagct taacgttggc ttgccacgca
     4321 ttacttgact gtaaaactct cactcttacc gaacttggcc gtaacctgcc aaccaaagcg
     4381 agaacaaaac ataacatcaa acgaatcgac cgattgttag gtaatcgtca cctccacaaa
     4441 gagcgactcg ctgtataccg ttggcatgct agctttatct gttcgggcaa tacgatgccc
     4501 attgtacttg ttgactggtc tgatattcgt gagcaaaaac gacttatggt attgcgagct
     4561 tcagtcgcac tacacggtcg ttctgttact ctttatgaga aagcgttccc gctttcagag
     4621 caatgttcaa agaaagctca tgaccaattt ctagccgacc ttgcgagcat tctaccgagt
     4681 aacaccacac cgctcattgt cagtgatgct ggctttaaag tgccatggta taaatccgtt
     4741 gagaagctgg gttggtactg gttaagtcga gtaagaggaa aagtacaata tgcagaccta
     4801 ggagcggaaa actggaaacc tatcagcaac ttacatgata tgtcatctag tcactcaaag
     4861 actttaggct ataagaggct gactaaaagc aatccaatct catgccaaat tctattgtat
     4921 aaatctcgct ctaaaggccg aaaaaatcag cgctcgacac ggactcattg tcaccacccg
     4981 tcacctaaaa tctactcagc gtcggcaaag gagccatggg ttctagcaac taacttacct
     5041 gttgaaattc gaacacccaa acaacttgtt aatatctatt cgaagcgaat gcagattgaa
     5101 gaaaccttcc gagacttgaa aagtcctgcc tacggactag gcctacgcca tagccgaacg
     5161 agcagctcag agcgttttga tatcatgctg ctaatcgccc tgatgcttca actaacatgt
     5221 tggcttgcgg gcgttcatgc tcagaaacaa ggttgggaca agcacttcca ggctaacaca
     5281 gtcagaaatc gaaacgtact ctcaacagtt cgcttaggca tggaagtttt gcggcattct
     5341 ggctacacaa taacaaggga agacttactc gtggctgcaa ccctactagc tcaaaattta
     5401 ttcacacatg gttacgcttt ggggaaatta tgaggggatc tctcagcgcc gggcgttttt
     5461 tattggtgag aatccaagcc tcgagctgtc agaccaagtt tactcatata tactttagat
     5521 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5761 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctgtg
     6241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6301 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6361 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6421 ctgcataacg cgaagtaatc ttttcggttt taaagaaaaa gggcagggtg gtgacacctt
     6481 gccctttttt gccggactgc agcggccgcg tatcccatca tcagacatac tcgatgagtt
     6541 aatacaaaaa aaaaaattgc aggtgcttta taaagtaaaa taacatcgtg gaatgcaagt
     6601 tattttttag tcatccacaa ttgctgcagt ggtattatga ggcaggattt ttttgacacc
     6661 acaattatgg tattttcgtg agcaaaccca acttttggta tatgtctaga tgcagagagt
     6721 gccgtaatcc tttttcttgt gacctttttc ttagcacact actttttgat gtggaaatat
     6781 aagttatgca agagaaaaaa cagaagggct ttacaaaatt atcctccttc cgattctgaa
     6841 ggtgggctct ctaaagtatc taaaatcgta ggggagtact gctactacca tcactgacat
     6901 ttggggctta ttcttttctg aaaattcaag atttttcaga cacgaccaaa caattagtac
     6961 gcaccgacta ctgcagttat ttatctgtca agccacagaa gtttctgcat gcactagt
//
