LOCUS       sequence-322120-        4293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reversed sequences of the NOS terminator, Firefly Luciferase CDS and
            5' UTR of 35S promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  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 4293)
  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 TNos:Luc:5'UTR (GB1499)
FEATURES             Location/Qualifiers
     source          1..4293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      254..348
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      531..1119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1020..1039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1273..1290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1314..1371)
                     /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."
     terminator      complement(1633..1885)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1912..3564)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     3480..3498
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    3616..3637
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      3640..3683
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             join(3867..4293,1..233)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3933..3952)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttttcgtatc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
       61 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
      121 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
      181 gaatgcttaa tgaattacaa cagtactgcg acgagtggca gggcggggcg taatttgata
      241 tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
      301 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcctcgag
      361 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
      421 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
      481 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      541 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      601 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      661 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      721 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      781 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
      841 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
      901 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
      961 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1021 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1081 tttttgtgat gctcgtcagg ggggcggagc ctgtggaaaa acgccagcaa cgcggccttt
     1141 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1201 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1261 acgaccgagc gcagcgagtc agtgagcgag gaagcctgca taacgcgaag taatcttttc
     1321 ggttttaaag aaaaagggca gggtggtgac accttgccct tttttgccgg actgcagcgg
     1381 ccgctactag tcgcgatgag tggtctcagg agcgagtcgg tcccattatt gaagcattta
     1441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1561 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattagc
     1621 ttggcaattc ccgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg
     1681 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa
     1741 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat
     1801 tcaacagaaa ttatatgata atcgtcgcaa gaccggcaac aggattcaat cttaagaaac
     1861 tttattgcca aatgtttgaa cgatcgggat cgaagatcca ttccaagctc attacacggc
     1921 gatctttccg cccttcttgg cctttatgag gatctctctg atttttcttg cgtcgagttt
     1981 tccggtaaga cctttcggta cttcgtccac aaacacaact cctccgcgca actttttcgc
     2041 ggttgttact tgactggcga cgtaatccac gatctctttt tccgtcatcg tctttccgtg
     2101 ctccaaaaca acaacggcgg cgggaagttc accggcgtca tcgtcgggaa gacctgcgac
     2161 acctgcgtcg aagatgttgg ggtgttggag caagatggat tccaattcag cgggagccac
     2221 ctgatagcct ttgtacttaa tcagagactt caggcggtca acgatgaaga agtgttcgtc
     2281 ttcgtcccag taagctatgt ctccagaatg tagccatcca tccttgtcaa tcaaggcgtt
     2341 ggtcgcttcc ggattgttta cataaccgga cataatcata ggacctctca cacacagttc
     2401 gcctctttga ttaacgccca gcgttttccc ggtatccaga tccacaacct tcgcttcaaa
     2461 aaatggaaca actttaccga ccgcgcccgg tttatcatcc ccctcgggtg taatcagaat
     2521 agctgatgta gtctcagtga gcccatatcc ttgcctgata cctggcagat ggaacctctt
     2581 ggcaaccgct tccccgactt ccttagagag gggagcgcca ccagaagcaa tttcgtgtaa
     2641 attagataaa tcgtatttgt caatcagagt gcttttggcg aagaaggaga atagggttgg
     2701 caccagcagc gcactttgaa tcttgtaatc ctgaaggctc ctcagaaaca gctcttcttc
     2761 aaatctatac attaagacga ctcgaaatcc acatatcaaa tatccgagtg tagtaaacat
     2821 tccaaaaccg tgatggaatg gaacaacact taaaatcgca gtatccggaa tgatttgatt
     2881 gccaaaaata ggatctctgg catgcgagaa tctcacgcag gcagttctat gaggcagagc
     2941 gacaccttta ggcagaccag tagatccaga ggagttcatg atcagtgcaa ttgtcttgtc
     3001 cctatcgaag gactctggca caaaatcgta ttcattaaaa ccgggaggta gatgagatgt
     3061 gacgaacgtg tacatcgact gaaatccctg gtaatccgtt ttagaatcca tgataataat
     3121 tttttggatg attgggagct ttttttgcac gttcaaaatt ttttgcaacc cctttttgga
     3181 aacgaacacc acggtaggct gcgaaatgcc catactgttg agcaattcac gttcattata
     3241 aatgtcgttc gcgggcgcaa ctgcaactcc gataaataac gcgcccaaca ccggcataaa
     3301 gaattgaaga gagttttcac tgcatacgac gattctgtga tttgtattca gcccatatcg
     3361 tttcatagct tctgccaacc gaacggacat ttcgaagtac tcagcgtaag tgatgtccac
     3421 ctcgatatgt gcatctgtaa aagcaattgt tccaggaacc agggcgtatc tcttcatagc
     3481 cttatgcagt tgctctccag cggttccatc ttccagcgga tagaatggcg ccgggccttt
     3541 ctttatgttt ttggcgtctt ccatttcgac tagaatagta aattgtaatg ttgtttctat
     3601 cgagaccact catcgctcta gaagcggccg cgaattccag aaatcatcct tagcgaaagc
     3661 taaggatttt ttttatctga aattctgcct cgtgatacgc ctatttttat aggttaatgt
     3721 catgataata atggtttctt agacgtcagg tggcactcga gttgatcggg cacgtaagag
     3781 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     3841 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     3901 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     3961 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     4021 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     4081 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     4141 acaccgtttt ccatgagcaa actgaaacgt tttcatcact ctggagtgaa taccacgacg
     4201 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     4261 cctatttccc taaagggttt attgagaata tgt
//
