LOCUS       sequence-204882-        4808 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway entry clone generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4808)
  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 4808)
  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: pJOG130
FEATURES             Location/Qualifiers
     source          1..4808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(706..711,712..729,730..736)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             790..1449
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(856..875)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1791..2096
                     /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     1975..1994
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2157..2256)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2273..2292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2274..2292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2297..2313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2297..2313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     mobile_element  2355..3122
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     primer_bind     complement(3191..3210)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3202..4011
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3269..3288)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4158..4746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4647..4666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgcaa tgtgagacca ttaggcaccc caggctttac actttatgct
      721 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
      781 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
      841 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
      901 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
      961 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     1021 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1081 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1141 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1201 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     1261 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1321 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1381 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1441 ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     1501 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     1561 ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     1621 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     1681 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     1741 gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta
     1801 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     1861 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     1921 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     1981 tgaccaccga tatggccagt gtgccggttt ccgttatcgg ggaagaagtg gctgatctca
     2041 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     2101 caggctccct tatacacagc cagtctgcag gtcgacggtc tcagcttggc gcgccgaccc
     2161 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2221 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2281 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2341 ctgatgttac attgggtaat gactccaact tattgatagt gttttatgtt cagataatgc
     2401 ccgatgactt tgtcatgcag ctccaccgat tttgagaacg acagcgactt ccgtcccagc
     2461 cgtgccaggt gctgcctcag attcaggtta tgccgctcaa ttcgctgcgt atatcgcttg
     2521 ctgattacgt gcagctttcc cttcaggcgg gattcataca gcggccagcc atccgtcatc
     2581 catatcacca cgtcaaaggg tgacagcagg ctcataagac gccccagcgt cgccatagtg
     2641 cgttcaccga atacgtgcgc aacaaccgtc ttccggagac tgtcatacgc gtaaaacagc
     2701 cagcgctggc gcgatttagc cccgacatag ccccactgtt cgtccatttc cgcgcagacg
     2761 atgacgtcac tgcccggctg tatgcgcgag gttaccgact gcggcctgag ttttttaagt
     2821 gacgtaaaat cgtgttgagg ccaacgccca taatgcgggc tgttgcccgg catccaacgc
     2881 cattcatggc catatcaatg attttctggt gcgtaccggg ttgagaagcg gtgtaagtga
     2941 actgcagttg ccatgtttta cggcagtgag agcagagata gcgctgatgt ccggcggtgc
     3001 ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagctg atagaaacag
     3061 aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca
     3121 ccttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     3181 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     3241 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     3301 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     3361 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     3421 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     3481 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     3541 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3601 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     3661 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3721 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3781 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3841 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3901 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3961 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4021 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4081 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     4141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4801 cacatgtt
//
