LOCUS       sequence-207475-        6393 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 6393)
  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 6393)
  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: pCK013
FEATURES             Location/Qualifiers
     source          1..6393
                     /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             3195..3224
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3231..3260
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3834..4496)
                     /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      4497..5268
                     /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 tctcagctta acaaaagttg atttctgaag
     3181 aagatttgaa cggtgaacaa aagctaatct ccgaggaaga cttgaacggt gaacaaaaat
     3241 taatctcaga agaagacttg aactgaagct ttaagcgtta atattttgtt aaaattcgcg
     3301 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgtactgcg atgagtggca
     3361 gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg gttgctacgc
     3421 ctgaataagt gataataagc ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc
     3481 gtcggttcag ggcagggtcg ttaaatagcc gcttatgtct attgctggtt taccggttta
     3541 ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag
     3601 gctctccccg tggaggtaat aattgacgat atgatcattt attctgcctc ccagagcctg
     3661 ataaaaacgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     3721 gactccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgaggcggc
     3781 tacagccgat agtctggaac agcgcactta cgggttgctg cgcaacccaa gtgctaccgg
     3841 cgcggcagcg tgacccgtgt cggcggctcc aacggctcgc catcgtccag aaaacacggc
     3901 tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc attcctccgt ttcggtcaag
     3961 gctggcaggt ctggttccat gcccggaatg ccgggctggc tgggcggctc ctcgccgggg
     4021 ccggtcggta gttgctgctc gcccggatac agggtcggga tgcggcgcag gtcgccatgc
     4081 cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca cggcggcact gaacaccgac
     4141 aggcgcaact ggtcgcgggg ctggccccac gccacgcggt cattgaccac gtaggccgac
     4201 acggtgccgg ggccgttgag cttcacgacg gagatccagc gctcggccac caagtccttg
     4261 actgcgtatt ggaccgtccg caaagaacgt ccgatgagct tggaaagtgt cttctggctg
     4321 accaccacgg cgttctggtg gcccatctgc gccacgaggt gatgcagcag cattgccgcc
     4381 gtgggtttcc tcgcaataag cccggcccac gcctcatgcg ctttgcgttc cgtttgcacc
     4441 cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc tggactgcgt ggccatgctt
     4501 atctccatgc ggtaggggtg ccgcacggtt gcggcaccat gcgcaatcag ctgcaacttt
     4561 tcggcagcgc gacaacaatt atgcgttgcg taaaagtggc agtcaattac agattttctt
     4621 taacctacgc aatgagctat tgcggggggt gccgcaatga gctgttgcgt accccccttt
     4681 tttaagttgt tgatttttaa gtctttcgca tttcgcccta tatctagttc tttggtgccc
     4741 aaagaagggc acccctgcgg ggttccccca cgccttcggc gcggctcccc ctccggcaaa
     4801 aagtggcccc tccggggctt gttgatcgac tgcgcggcct tcggccttgc ccaaggtggc
     4861 gctgccccct tggaaccccc gcactcgccg ccgtgaggct cggggggcag gcgggcgggc
     4921 ttcgcccttc gactgccccc actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca
     4981 agccgctgcg cggtcgctgc gcgagccttg acccgccttc cacttggtgt ccaaccggca
     5041 agcgaagcgc gcaggccgca ggccggaggc ttttccccag agaaaattaa aaaaattgat
     5101 ggggcaaggc cgcaggccgc gcagttggag ccggtgggta tgtggtcgaa ggctgggtag
     5161 ccggtgggca atccctgtgg tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc
     5221 cagcagggtt gtccacgggc cgagcgaagc gagccagccg gtggccgctc gcggccatcg
     5281 tccacatatc cacgggctgg caagggagcg cagcgaccgc gcagggcgaa gcccggagag
     5341 caagcccgta gggcgccgca gccgccgtag gcggtcacga ctttgcgaag caaagtctag
     5401 tgagtatact caagcattga gtggcccgcc ggaggcaccg ccttgcgctg cccccgtcga
     5461 gccggttgga caccaaaagg gaggggcagg catggcggca tacgcgatca tgcgatgcaa
     5521 gaagctggcg aaaatgggca acgtggcggc cagtctcaag cacgcctacc gcgagcgcga
     5581 gacgcccaac gctgacgcca gcaggacgcc agagaacgag cactgggcgg ccagcagcac
     5641 cgatgaagcg atgggccgac tgcgcgagtt gctgccagag aagcggcgca aggacgctgt
     5701 gttggcggtc gagtacgtca tgacggccag cccggaatgg tggaagtcgg ccagccaaga
     5761 acagcaggcg gcgttcttcg agaaggcgca caagtggctg gcggacaagt acggggcgga
     5821 tcgcatcgtg acggccagca tccaccgtga cgaaaccagc ccgcacatga ccgcgttcgt
     5881 ggtgccgctg acgcaggacg gcaggctgtc ggccaaggag ttcatcggca acaaagcgca
     5941 gatgacccgc gaccagacca cgtttgcggc cgctgtggcc gatctagggc tgcaacgggg
     6001 catcgagggc agcaaggcac gtcacacgcg cattcaggcg ttctacgagg ccctggagcg
     6061 gccaccagtg ggccacgtca ccatcagccc gcaagcggtc gagccacgcg cctatgcacc
     6121 gcagggattg gccgaaaagc tgggaatctc aaagcgcgtt gagacgccgg aagccgtggc
     6181 cgaccggctg acaaaagcgg ttcggcaggg gtatgagcct gccctacagg ccgccgcagg
     6241 agcgcgtgag atgcgcaaga aggccgatca agcccaagag acggcccgag atcttcggga
     6301 gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc aggccaaggc
     6361 cgccgcgatc atcaaggccg tgggcgaaaa gct
//
