LOCUS       sequence-495508-        3355 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Level 0 Golden Gate Promoter + 5' UTR acceptor with AmilCP negative 
            selection cassette GGAG - CCAT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3355)
  TITLE     Plant Golden-gate Promoters, Terminators and Level0/Level1 cloning 
            acceptors.
REFERENCE   2  (bases 1 to 3355)
  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: pICSL01008_CP
FEATURES             Location/Qualifiers
     source          1..3355
                     /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"
     protein_bind    960..981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(996..1001,1002..1019,1020..1026)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1034..1050
                     /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             1068..1079
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1798..1869
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1885..1912
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1982..2000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2068..2172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2394..3185
                     /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 tcactctgtg gtctcaggag
      841 ttgtcttcgc agctggcacc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      901 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      961 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     1021 atgttgtgtg gaattgtgag cggataacaa tttcacacat actagagaaa gaggagaaat
     1081 actagaatga gcgtgatcgc taagcagatg acctacaagg tgtacatgag cggaactgtg
     1141 aacggtcact acttcgaggt tgagggtgat ggtaagggta agccttatga gggtgagcag
     1201 actgtgaagc ttactgtgac taagggtggt cctcttcctt tcgcttggga tattttgtct
     1261 cctcagtgcc agtacggtag catccctttc actaagtacc ctgaggatat ccctgattac
     1321 gtgaagcagt ctttcccaga gggttacacc tgggagagga tcatgaattt cgaggatggt
     1381 gctgtgtgca ccgtgagcaa cgattcttct attcagggaa actgcttcat ctaccacgtg
     1441 aagttctccg gtctgaactt ccctcctaat ggtcctgtga tgcagaagaa aactcagggt
     1501 tgggagccta acaccgagag gctttttgct agggatggta tgctgctggg taacaacttc
     1561 atggctctga agcttgaggg tggtggacat tacctttgcg agttcaagac cacttacaag
     1621 gctaagaaac ctgtgaagat gcctggttac cactatgtgg atagaaagct ggatgtgacc
     1681 aaccacaaca aggattacac cagcgttgag cagtgcgaga tctctattgc tagaaagcct
     1741 gtggtggctt aggcttcgct gatagtgcta gtgtagatcg ctactagagc caggcatcaa
     1801 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1861 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttataac
     1921 agacaagctg tgacgaagac aaccattgag accacgaagt ggctcttcag tggacgaaag
     1981 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2041 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2101 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2161 aaaaggaaga gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     2221 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttttggg gtacagtcta
     2281 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     2341 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac atcatgaggg
     2401 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
     2461 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
     2521 agccacacag cgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     2581 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
     2641 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     2701 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
     2761 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     2821 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
     2881 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     2941 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     3001 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     3061 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     3121 cagatcagtt ggaagaattt gtccattacg taaaaggcga gatcaccaag gtagtcggca
     3181 aataactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3241 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3301 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
