LOCUS       sequence-207479-        6297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial translocation fo proteins into plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  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 6297)
  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"; journalName: "PLoS One.";
            date: "2018-05-30"; volume: "13(5):e0197185. doi"; pages: " 
            10.1371/journal.pone.0197185. eCollection 2018"
COMMENT     Sequence Label: pCK015
FEATURES             Location/Qualifiers
     source          1..6297
                     /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             3099..3128
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3135..3164
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3738..4400)
                     /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      4401..5172
                     /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 tcggtctcag
     3061 cttaacaaaa gttgatttct gaagaagatt tgaacggtga acaaaagcta atctccgagg
     3121 aagacttgaa cggtgaacaa aaattaatct cagaagaaga cttgaactga agctttaagc
     3181 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3241 taggccgtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     3301 cccttaaacg cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     3361 ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat
     3421 gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca
     3481 aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc
     3541 atttattctg cctcccagag cctgataaaa acggtgaatc cgttagcgag gtgccgccgg
     3601 cttccattca ggtcgaggtg gcccgactcc atgcaccgcg acgcaacgcg gggaggcaga
     3661 caaggtatag ggcggcgagg cggctacagc cgatagtctg gaacagcgca cttacgggtt
     3721 gctgcgcaac ccaagtgcta ccggcgcggc agcgtgaccc gtgtcggcgg ctccaacggc
     3781 tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg cccgcgccgt
     3841 tcccattcct ccgtttcggt caaggctggc aggtctggtt ccatgcccgg aatgccgggc
     3901 tggctgggcg gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg atacagggtc
     3961 gggatgcggc gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc gtgatcaacc
     4021 accacggcgg cactgaacac cgacaggcgc aactggtcgc ggggctggcc ccacgccacg
     4081 cggtcattga ccacgtaggc cgacacggtg ccggggccgt tgagcttcac gacggagatc
     4141 cagcgctcgg ccaccaagtc cttgactgcg tattggaccg tccgcaaaga acgtccgatg
     4201 agcttggaaa gtgtcttctg gctgaccacc acggcgttct ggtggcccat ctgcgccacg
     4261 aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca
     4321 tgcgctttgc gttccgtttg cacccagtga ccgggcttgt tcttggcttg aatgccgatt
     4381 tctctggact gcgtggccat gcttatctcc atgcggtagg ggtgccgcac ggttgcggca
     4441 ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag
     4501 tggcagtcaa ttacagattt tctttaacct acgcaatgag ctattgcggg gggtgccgca
     4561 atgagctgtt gcgtaccccc cttttttaag ttgttgattt ttaagtcttt cgcatttcgc
     4621 cctatatcta gttctttggt gcccaaagaa gggcacccct gcggggttcc cccacgcctt
     4681 cggcgcggct ccccctccgg caaaaagtgg cccctccggg gcttgttgat cgactgcgcg
     4741 gccttcggcc ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga
     4801 ggctcggggg gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc ataggcttgg
     4861 gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc
     4921 cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc
     4981 ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg
     5041 ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag
     5101 gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca
     5161 gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga
     5221 ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc
     5281 acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc
     5341 accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc
     5401 ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct
     5461 caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa
     5521 cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc
     5581 agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga
     5641 atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg
     5701 gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac
     5761 cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa
     5821 ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt
     5881 ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca
     5941 ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc
     6001 ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg
     6061 cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga
     6121 gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca
     6181 agagacggcc cgagatcttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt
     6241 gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagct
//
