LOCUS       sequence-210238-        6394 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 6394)
  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 6394)
  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: pCK014
FEATURES             Location/Qualifiers
     source          1..6394
                     /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        1708..1810
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1811..2494
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(1877..1896)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2814..3119
                     /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     2998..3017
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3160..3189
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3196..3225
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3232..3261
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3835..4497)
                     /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      4498..5269
                     /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 aattatgact cgaatttcaa ccagttcagt aaattccagc
     1321 ttcagttact cggcccctgc agaggaagcg caaaaccgcg tttcctcagc ccccaccaac
     1381 tcctctccac ctggcaccac aaccgctgtc gcccaggcgt cagaggggca aaaaaagccg
     1441 ggggcggtat taagcatgca tgcacagcga ctgcgccaat tgtgtgagag ttcatctcac
     1501 cagtaccggc agaacgcgct attagctaaa tcttttgatg ctcaacgcct agacattaac
     1561 actcaagcgg gcccttccaa tagccctcac ctgaacgctc tcaacaagct ccaacaacga
     1621 gatttcaaac ctgcggctgg cgggctagaa atcccattta cacccaaatc cctattgggt
     1681 ggaggtgcaa tgagagacct gagacgttga tcggcacgta agaggttcca actttcacca
     1741 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     1801 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     1861 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1921 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1981 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
     2041 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     2101 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     2161 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     2221 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     2281 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     2341 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     2401 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     2461 ggggcgtaaa cgccgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg
     2521 cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa
     2581 agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc
     2641 tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc
     2701 ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc
     2761 ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt
     2821 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     2881 atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     2941 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     3001 tgatgaccac cgatatggcc agtgtgccgg tttccgttat cggggaagaa gtggctgatc
     3061 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     3121 tgtcaggctc ccttatacac agccagtcgg tctcattcgg aacaaaagtt gatttctgaa
     3181 gaagatttga acggtgaaca aaagctaatc tccgaggaag acttgaacgg tgaacaaaaa
     3241 ttaatctcag aagaagactt gaactgaagc tttaagcgtt aatattttgt taaaattcgc
     3301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgtactgc gatgagtggc
     3361 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     3421 cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat tcgacccggt
     3481 cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt ttaccggttt
     3541 attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc agtttgctca
     3601 ggctctcccc gtggaggtaa taattgacga tatgatcatt tattctgcct cccagagcct
     3661 gataaaaacg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     3721 cgactccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgaggcgg
     3781 ctacagccga tagtctggaa cagcgcactt acgggttgct gcgcaaccca agtgctaccg
     3841 gcgcggcagc gtgacccgtg tcggcggctc caacggctcg ccatcgtcca gaaaacacgg
     3901 ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc cattcctccg tttcggtcaa
     3961 ggctggcagg tctggttcca tgcccggaat gccgggctgg ctgggcggct cctcgccggg
     4021 gccggtcggt agttgctgct cgcccggata cagggtcggg atgcggcgca ggtcgccatg
     4081 ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc acggcggcac tgaacaccga
     4141 caggcgcaac tggtcgcggg gctggcccca cgccacgcgg tcattgacca cgtaggccga
     4201 cacggtgccg gggccgttga gcttcacgac ggagatccag cgctcggcca ccaagtcctt
     4261 gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc ttggaaagtg tcttctggct
     4321 gaccaccacg gcgttctggt ggcccatctg cgccacgagg tgatgcagca gcattgccgc
     4381 cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc gctttgcgtt ccgtttgcac
     4441 ccagtgaccg ggcttgttct tggcttgaat gccgatttct ctggactgcg tggccatgct
     4501 tatctccatg cggtaggggt gccgcacggt tgcggcacca tgcgcaatca gctgcaactt
     4561 ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta cagattttct
     4621 ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg tacccccctt
     4681 ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt ctttggtgcc
     4741 caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc cctccggcaa
     4801 aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg
     4861 cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg
     4921 cttcgccctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc
     4981 aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg tccaaccggc
     5041 aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta aaaaaattga
     5101 tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga aggctgggta
     5161 gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt
     5221 ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct cgcggccatc
     5281 gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga agcccggaga
     5341 gcaagcccgt agggcgccgc agccgccgta ggcggtcacg actttgcgaa gcaaagtcta
     5401 gtgagtatac tcaagcattg agtggcccgc cggaggcacc gccttgcgct gcccccgtcg
     5461 agccggttgg acaccaaaag ggaggggcag gcatggcggc atacgcgatc atgcgatgca
     5521 agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac cgcgagcgcg
     5581 agacgcccaa cgctgacgcc agcaggacgc cagagaacga gcactgggcg gccagcagca
     5641 ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc aaggacgctg
     5701 tgttggcggt cgagtacgtc atgacggcca gcccggaatg gtggaagtcg gccagccaag
     5761 aacagcaggc ggcgttcttc gagaaggcgc acaagtggct ggcggacaag tacggggcgg
     5821 atcgcatcgt gacggccagc atccaccgtg acgaaaccag cccgcacatg accgcgttcg
     5881 tggtgccgct gacgcaggac ggcaggctgt cggccaagga gttcatcggc aacaaagcgc
     5941 agatgacccg cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg ctgcaacggg
     6001 gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc gttctacgag gccctggagc
     6061 ggccaccagt gggccacgtc accatcagcc cgcaagcggt cgagccacgc gcctatgcac
     6121 cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg gaagccgtgg
     6181 ccgaccggct gacaaaagcg gttcggcagg ggtgtgagcc tgccctacag gccgccgcag
     6241 gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga gacggcccga gatcttcggg
     6301 agcgcctgaa gcccgttctg gacgccctgg ggccgttgaa tcgggatatg caggccaagg
     6361 ccgccgcgat catcaaggcc gtgggcgaaa agct
//
