LOCUS       sequence-106088-        3138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Accepts DNA sequences via BpiI cloning sites, resulting in Level 0 
            standard parts for Golden Gate cloning. Parts can be released with a
            user-defined 4-base-pair, 5 prime overhangs using BsaI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3138)
  AUTHORS   Patron NJ, Orzaez D, Marillonnet S, Warzecha H, Matthewman C, Youles
            M, Raitskin O, Leveau A, Farre G, Rogers C, Smith A, Hibberd J, Webb
            AA, Locke J, Schornack S, Ajioka J, Baulcombe DC, Zipfel C, Kamoun 
            S, Jones JD, Kuhn H, Robatzek S, Van Esse HP, Sanders D, Oldroyd G, 
            Martin C, Field R, O'Connor S, Fox S, Wulff B, Miller B, Breakspear 
            A, Radhakrishnan G, Delaux PM, Loque D, Granell A, Tissier A, Shih 
            P, Brutnell TP, Quick WP, Rischer H, Fraser PD, Aharoni A, Raines C,
            South PF, Ane JM, Hamberger BR, Langdale J, Stougaard J, Bouwmeester
            H, Udvardi M, Murray JA, Ntoukakis V, Schafer P, Denby K, Edwards 
            KJ, Osbourn A, Haseloff J
  TITLE     Standards for plant synthetic biology: a common syntax for exchange 
            of DNA parts.
  JOURNAL   New Phytol. 2015 Jul 14. doi: 10.1111/nph.13532.
  PUBMED    26171760
REFERENCE   2  (bases 1 to 3138)
  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"; journalName: "New Phytol.
            2015 Jul 14. doi: 10.1111/nph.13532."
COMMENT     Sequence Label: pUAP1
FEATURES             Location/Qualifiers
     source          1..3138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      7..64
                     /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."
     primer_bind     complement(88..105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(259..847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(339..358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1030..1124)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1145..1804)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1719..1738
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1966..1984
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1988..2031)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    2155..2176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2191..2196,2197..2214,2215..2221)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2229..2245
                     /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             2263..2274
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2281..2958
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3003..3074
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3090..3117
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
       61 aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
      121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      241 ggccgcgttg ctggcgtttt tccacaggct ccgcccccct gacgagcatc acaaaaatcg
      301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      781 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      841 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      901 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      961 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagctc
     1021 gaggcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     1081 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat
     1141 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     1201 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     1261 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     1321 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     1381 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     1441 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     1501 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     1561 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     1621 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     1681 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     1741 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     1801 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     1861 atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaac tcgagtgcca
     1921 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1981 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
     2041 gtctcattgt cttccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2101 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2161 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2221 gtgtggaatt gtgagcggat aacaatttca cacatactag agaaagagga gaaatactag
     2281 atggcttcct ccgaggacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt
     2341 tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt
     2401 acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc
     2461 ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg
     2521 gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgag
     2581 gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac
     2641 aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc
     2701 atgggttggg aagcttccac cgaacgtatg tacccggagg acggtgctct gaaaggtgaa
     2761 atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc
     2821 tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac
     2881 atcacctccc acaacgagga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt
     2941 cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact agagccaggc
     3001 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3061 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     3121 atagaagaca acgagacc
//
