LOCUS       sequence-322121-        3620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  35S promoter sequence and the Mtb terminator flanked by two opposing
            PhiC31 site-specific recombination sites (attP and attB).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3620)
  AUTHORS   Bernabe-Orts JM, Quijano-Rubio A, Vazquez-Vilar M, Mancheno-Bonillo 
            J, Moles-Casas V, Selma S, Gianoglio S, Granell A, Orzaez D
  TITLE     A memory switch for plant synthetic biology based on the phage 
            varphiC31 integration system.
  JOURNAL   Nucleic Acids Res. 2020 Apr 6;48(6):3379-3394. doi: 
            10.1093/nar/gkaa104.
  PUBMED    32083668
REFERENCE   2  (bases 1 to 3620)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2020-04-6- 6"; volume: "48"; issue: "6";
            pages: "3379-3394"
COMMENT     Sequence Label: pUPD2 PhiC31 attP:TMtb:P35S:attB (GB1494)
FEATURES             Location/Qualifiers
     source          1..3620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     365..384
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     612..630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(634..677)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    680..701
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        1775..2119
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2063..2083
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    2190..2223
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     terminator      2273..2330
                     /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(2354..2371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2525..3113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2605..2624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3296..3390)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3411..3620,1..450))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ccagggattg gctgatacga aaaacatatt ctcaataaac cctttaggga aataggccag
       61 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
      121 gtggtattca ctccagagtg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
      181 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg
      241 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
      301 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
      361 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
      421 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
      481 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
      541 tacgtgcccg atcaactcga gtgccacctg acgtctaaga aaccattatt atcatgacat
      601 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
      661 tcgctaagga tgatttctgg aattcgcggc cgcttctaga gcgatgagtg gtctcgctat
      721 agtagtgccc caactggggt aacctttgag ttctctcagt tgggggcgta gtcgcaaaaa
      781 ctatatgctc tccaaccaca cccaatggaa ttaggtttta ccaaaaccta aaagatttac
      841 tatagttttg cagcctagaa acaggaaaac attatcaagg aagtacttga attagttgaa
      901 accacaacac atatccaaac ttatatgtta aaattaacag atgcagagat attcatgttt
      961 acacaaacca ttttactggt tacttacaca taaagactcc attactatat ttcttattac
     1021 agggaaaaaa gaacacattt ttttccctgt tttcaacaga cagacacact aaaacactag
     1081 acacacacat tttttcaact atttttcttc agccatagaa ggctaacacc atgatctctg
     1141 ctctgtttca actatatttt cgtactagag ccaagctgat ctcctttgcc ccggagatca
     1201 ccatggacga ctttctctat ctctacgatc taggaagaaa gttcgacgga gaaggtgacg
     1261 ataccatgtt caccaccgat aatgagaaga ttagcctctt caatttcaga aagaatgctg
     1321 acccacagat ggttagagag gcctacgcgg caggtctgat caagacgatc tacccgagta
     1381 ataatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca
     1441 ggactaactg catcaagaac acagagaaag atatatttct caagatcaga agtactattc
     1501 cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag attggagtct
     1561 ctaagaaagt agttcctact gaatcaaagg ccatggagtc aaaaattcag atcgaggatc
     1621 taacagaact cgccgtgaag actggcgaac agttcataca gagtctttta cgactcaatg
     1681 acaagaagaa aatcttcgtc aacatggtgg agcacgacac tctcgtctac tccaagaata
     1741 tcaaagatac agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaatat
     1801 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag
     1861 aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag
     1921 atgcccctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
     1981 aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg
     2041 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
     2101 catttcattt ggagaggact ccggtatttt tacaacaatt accacaacaa aacaaacaac
     2161 aaacaacatt acaatttact agtggagtac gcgcccgggg agcccaaggg cacgccctgg
     2221 cacccgcacc gaccttgaga ccactcatcg cgactagtag cggccgctgc agtccggcaa
     2281 aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg
     2341 ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2521 cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2941 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctcgagg cttggattct
     3301 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     3361 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
     3421 ccctgccact cgtcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     3481 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     3541 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     3601 tcaaaactgg tgaaactcac
//
