LOCUS       sequence-495507-        3350 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Level 0 Golden Gate C-terminal tag acceptor with AmilCP negative 
            selection cassette TTCG - GCTT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3350)
  TITLE     Plant Golden-gate Promoters, Terminators and Level0/Level1 cloning 
            acceptors.
REFERENCE   2  (bases 1 to 3350)
  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: pICSL01003_CP
FEATURES             Location/Qualifiers
     source          1..3350
                     /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    958..979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(994..999,1000..1017,1018..1024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1032..1048
                     /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             1066..1077
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1796..1867
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1883..1910
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1977..1995)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2063..2167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2389..3180
                     /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 tcactgtggt ctcattcgtt
      841 gtcttcgcag ctggcaccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      901 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      961 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1021 gttgtgtgga attgtgagcg gataacaatt tcacacatac tagagaaaga ggagaaatac
     1081 tagaatgagc gtgatcgcta agcagatgac ctacaaggtg tacatgagcg gaactgtgaa
     1141 cggtcactac ttcgaggttg agggtgatgg taagggtaag ccttatgagg gtgagcagac
     1201 tgtgaagctt actgtgacta agggtggtcc tcttcctttc gcttgggata ttttgtctcc
     1261 tcagtgccag tacggtagca tccctttcac taagtaccct gaggatatcc ctgattacgt
     1321 gaagcagtct ttcccagagg gttacacctg ggagaggatc atgaatttcg aggatggtgc
     1381 tgtgtgcacc gtgagcaacg attcttctat tcagggaaac tgcttcatct accacgtgaa
     1441 gttctccggt ctgaacttcc ctcctaatgg tcctgtgatg cagaagaaaa ctcagggttg
     1501 ggagcctaac accgagaggc tttttgctag ggatggtatg ctgctgggta acaacttcat
     1561 ggctctgaag cttgagggtg gtggacatta cctttgcgag ttcaagacca cttacaaggc
     1621 taagaaacct gtgaagatgc ctggttacca ctatgtggat agaaagctgg atgtgaccaa
     1681 ccacaacaag gattacacca gcgttgagca gtgcgagatc tctattgcta gaaagcctgt
     1741 ggtggcttag gcttcgctga tagtgctagt gtagatcgct actagagcca ggcatcaaat
     1801 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1861 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataacag
     1921 acaagctgtg acgaagacaa gctttgagac cacgtggctc ttcagtggac gaaagggcct
     1981 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     2041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2161 gaagagtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     2221 cgcagtggcg gttttcatgg cttgttatga ctgttttttt ttggggtaca gtctatgcct
     2281 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     2341 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacatcat gagggaagcg
     2401 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     2461 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     2521 cacagcgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     2581 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     2641 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     2701 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     2761 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     2821 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     2881 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     2941 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     3001 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     3061 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     3121 cagttggaag aatttgtcca ttacgtaaaa ggcgagatca ccaaggtagt cggcaaataa
     3181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
