LOCUS       sequence-495506-        3352 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Level 0 Golden Gate N-terminal tag acceptor with AmilCP negative 
            selection cassette CCAT - AATG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3352)
  TITLE     Plant Golden-gate Promoters, Terminators and Level0/Level1 cloning 
            acceptors.
REFERENCE   2  (bases 1 to 3352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pICSL01002_CP
FEATURES             Location/Qualifiers
     source          1..3352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(870..897)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(913..984)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1703..1714
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    1732..1748
                     /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(1756..1762,1763..1780,1781..1786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1801..1822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1979..1997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2065..2169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2391..3182
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactcgtgg tctcacattt
      841 tgtcttcgtc acagcttgtc tgttataaac gcagaaaggc ccacccgaag gtgagccagt
      901 gtgactctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
      961 cgactgagcc tttcgtttta tttgatgcct ggctctagta gcgatctaca ctagcactat
     1021 cagcgaagcc taagccacca caggctttct agcaatagag atctcgcact gctcaacgct
     1081 ggtgtaatcc ttgttgtggt tggtcacatc cagctttcta tccacatagt ggtaaccagg
     1141 catcttcaca ggtttcttag ccttgtaagt ggtcttgaac tcgcaaaggt aatgtccacc
     1201 accctcaagc ttcagagcca tgaagttgtt acccagcagc ataccatccc tagcaaaaag
     1261 cctctcggtg ttaggctccc aaccctgagt tttcttctgc atcacaggac cattaggagg
     1321 gaagttcaga ccggagaact tcacgtggta gatgaagcag tttccctgaa tagaagaatc
     1381 gttgctcacg gtgcacacag caccatcctc gaaattcatg atcctctccc aggtgtaacc
     1441 ctctgggaaa gactgcttca cgtaatcagg gatatcctca gggtacttag tgaaagggat
     1501 gctaccgtac tggcactgag gagacaaaat atcccaagcg aaaggaagag gaccaccctt
     1561 agtcacagta agcttcacag tctgctcacc ctcataaggc ttacccttac catcaccctc
     1621 aacctcgaag tagtgaccgt tcacagttcc gctcatgtac accttgtagg tcatctgctt
     1681 agcgatcacg ctcattctag tatttctcct ctttctctag tatgtgtgaa attgttatcc
     1741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1921 tggtgccagc tgcgaagaca aatggtgaga ccacagtggc tcttcagtgg acgaaagggc
     1981 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2041 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2101 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2161 aggaagagta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     2221 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttttggggta cagtctatgc
     2281 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
     2341 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc atgagggaag
     2401 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc
     2461 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc
     2521 cacacagcga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc
     2581 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc
     2641 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta
     2701 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc
     2761 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg
     2821 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg
     2881 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc
     2941 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc
     3001 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca
     3061 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag
     3121 atcagttgga agaatttgtc cattacgtaa aaggcgagat caccaaggta gtcggcaaat
     3181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
