LOCUS       sequence-207470-        6298 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial translocation for proteins into plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6298)
  AUTHORS   Gantner J, Ordon J, Ilse T, Kretschmer C, Gruetzner R, Lofke C, 
            Dagdas Y, Burstenbinder K, Marillonnet S, Stuttmann J
  TITLE     Peripheral infrastructure vectors and an extended set of plant parts
            for the Modular Cloning system.
  JOURNAL   PLoS One. 2018 May 30;13(5):e0197185. doi: 
            10.1371/journal.pone.0197185. eCollection 2018.
  PUBMED    29847550
REFERENCE   2  (bases 1 to 6298)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2018-05-30"; volume: "13(5):e0197185. doi"; pages: " 
            10.1371/journal.pone.0197185. eCollection 2018"
COMMENT     Sequence Label: pCK016
FEATURES             Location/Qualifiers
     source          1..6298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(157..690)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(879..884,885..901,902..907))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1177..1198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1213..1218,1219..1236,1237..1243)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1251..1267
                     /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)."
     primer_bind     1256..1278
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        1612..1714
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1715..2398
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(1781..1800)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2718..3023
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2902..2921
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3064..3093
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3100..3129
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3136..3165
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3739..4401)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      4402..5173
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 gctgacggaa cagcgggaag tccagcgcca gaaacaggcc cagcgccagc aggaacgcgg
       61 gcgcgcacat ttccccgaaa agtgccacct gcggcgttgt gacaatttac cgaacaactc
      121 cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat
      181 atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc
      241 gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt
      301 gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctctccgg agactgcgag
      361 atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag
      421 agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag
      481 caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa
      541 ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct
      601 acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg
      661 ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
      721 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt
      781 cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc
      841 tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt
      901 atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtccatgg gcaaatatta
      961 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1021 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
     1081 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1141 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1201 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1261 ataacaattt cacacaggag aattatgaaa attgctccag ttgccataaa tcacagcccg
     1321 ctcagcaggg aggtcccgtc acacgcggca cccactcagg caaagcaaac caaccttcaa
     1381 tctgaagctg gcgatttaga tgcaagaaaa agtagcgctt caagcccgga aacccgcgca
     1441 ttactcgcta ctaagacagt actcgggaga cacaagatag aggttccggc ctttggaggg
     1501 tggttcaaaa agaaatcatc taagcacgag acgggcggtt caagtgccaa cgcagatagt
     1561 tcgagcgtgg cttccgattc caccggtggc tcaatgagag acctgagacg ttgatcggca
     1621 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     1681 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     1741 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     1801 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     1861 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     1921 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     1981 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     2041 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     2101 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     2161 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     2221 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     2281 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     2341 acagtactgc gatgagtggc agggggggcg taaacgccgc gtggatccgg cttactaaaa
     2401 gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat
     2461 atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag
     2521 ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt
     2581 aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa
     2641 tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga
     2701 gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc
     2761 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc
     2821 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc
     2881 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtttccg
     2941 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
     3001 acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag tcggtctcat
     3061 tcggaacaaa agttgatttc tgaagaagat ttgaacggtg aacaaaagct aatctccgag
     3121 gaagacttga acggtgaaca aaaattaatc tcagaagaag acttgaactg aagctttaag
     3181 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3241 ataggccgta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     3301 gcccttaaac gcctggttgc tacgcctgaa taagtgataa taagcggatg aatggcagaa
     3361 attcgaaagc aaattcgacc cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta
     3421 tgtctattgc tggtttaccg gtttattgac taccggaagc agtgtgaccg tgtgcttctc
     3481 aaatgcctga ggccagtttg ctcaggctct ccccgtggag gtaataattg acgatatgat
     3541 catttattct gcctcccaga gcctgataaa aacggtgaat ccgttagcga ggtgccgccg
     3601 gcttccattc aggtcgaggt ggcccgactc catgcaccgc gacgcaacgc ggggaggcag
     3661 acaaggtata gggcggcgag gcggctacag ccgatagtct ggaacagcgc acttacgggt
     3721 tgctgcgcaa cccaagtgct accggcgcgg cagcgtgacc cgtgtcggcg gctccaacgg
     3781 ctcgccatcg tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg
     3841 ttcccattcc tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg
     3901 ctggctgggc ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt
     3961 cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac
     4021 caccacggcg gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac
     4081 gcggtcattg accacgtagg ccgacacggt gccggggccg ttgagcttca cgacggagat
     4141 ccagcgctcg gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat
     4201 gagcttggaa agtgtcttct ggctgaccac cacggcgttc tggtggccca tctgcgccac
     4261 gaggtgatgc agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc
     4321 atgcgctttg cgttccgttt gcacccagtg accgggcttg ttcttggctt gaatgccgat
     4381 ttctctggac tgcgtggcca tgcttatctc catgcggtag gggtgccgca cggttgcggc
     4441 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa
     4501 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc
     4561 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg
     4621 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct
     4681 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc
     4741 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg
     4801 aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg cccccactcg cataggcttg
     4861 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg
     4921 ccttccactt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc
     4981 cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt
     5041 gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca
     5101 ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccgagc gaagcgagcc
     5161 agccggtggc cgctcgcggc catcgtccac atatccacgg gctggcaagg gagcgcagcg
     5221 accgcgcagg gcgaagcccg gagagcaagc ccgtagggcg ccgcagccgc cgtaggcggt
     5281 cacgactttg cgaagcaaag tctagtgagt atactcaagc attgagtggc ccgccggagg
     5341 caccgccttg cgctgccccc gtcgagccgg ttggacacca aaagggaggg gcaggcatgg
     5401 cggcatacgc gatcatgcga tgcaagaagc tggcgaaaat gggcaacgtg gcggccagtc
     5461 tcaagcacgc ctaccgcgag cgcgagacgc ccaacgctga cgccagcagg acgccagaga
     5521 acgagcactg ggcggccagc agcaccgatg aagcgatggg ccgactgcgc gagttgctgc
     5581 cagagaagcg gcgcaaggac gctgtgttgg cggtcgagta cgtcatgacg gccagcccgg
     5641 aatggtggaa gtcggccagc caagaacagc aggcggcgtt cttcgagaag gcgcacaagt
     5701 ggctggcgga caagtacggg gcggatcgca tcgtgacggc cagcatccac cgtgacgaaa
     5761 ccagcccgca catgaccgcg ttcgtggtgc cgctgacgca ggacggcagg ctgtcggcca
     5821 aggagttcat cggcaacaaa gcgcagatga cccgcgacca gaccacgttt gcggccgctg
     5881 tggccgatct agggctgcaa cggggcatcg agggcagcaa ggcacgtcac acgcgcattc
     5941 aggcgttcta cgaggccctg gagcggccac cagtgggcca cgtcaccatc agcccgcaag
     6001 cggtcgagcc acgcgcctat gcaccgcagg gattggccga aaagctggga atctcaaagc
     6061 gcgttgagac gccggaagcc gtggccgacc ggctgacaaa agcggttcgg caggggtatg
     6121 agcctgccct acaggccgcc gcaggagcgc gtgagatgcg caagaaggcc gatcaagccc
     6181 aagagacggc ccgagatctt cgggagcgcc tgaagcccgt tctggacgcc ctggggccgt
     6241 tgaatcggga tatgcaggcc aaggccgccg cgatcatcaa ggccgtgggc gaaaagct
//
