LOCUS       sequence-204884-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4809)
  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 4809)
  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: pJOG131
FEATURES             Location/Qualifiers
     source          1..4809
                     /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(2158..2257)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2274..2293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2275..2293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2298..2314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2298..2314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     mobile_element  2356..3123
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     primer_bind     complement(3192..3211)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3203..4012
                     /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(3270..3289)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4159..4747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4648..4667
                     /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 tcattcgggg cgcgccgacc
     2161 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2221 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     2281 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     2341 tctgatgtta cattgggtaa tgactccaac ttattgatag tgttttatgt tcagataatg
     2401 cccgatgact ttgtcatgca gctccaccga ttttgagaac gacagcgact tccgtcccag
     2461 ccgtgccagg tgctgcctca gattcaggtt atgccgctca attcgctgcg tatatcgctt
     2521 gctgattacg tgcagctttc ccttcaggcg ggattcatac agcggccagc catccgtcat
     2581 ccatatcacc acgtcaaagg gtgacagcag gctcataaga cgccccagcg tcgccatagt
     2641 gcgttcaccg aatacgtgcg caacaaccgt cttccggaga ctgtcatacg cgtaaaacag
     2701 ccagcgctgg cgcgatttag ccccgacata gccccactgt tcgtccattt ccgcgcagac
     2761 gatgacgtca ctgcccggct gtatgcgcga ggttaccgac tgcggcctga gttttttaag
     2821 tgacgtaaaa tcgtgttgag gccaacgccc ataatgcggg ctgttgcccg gcatccaacg
     2881 ccattcatgg ccatatcaat gattttctgg tgcgtaccgg gttgagaagc ggtgtaagtg
     2941 aactgcagtt gccatgtttt acggcagtga gagcagagat agcgctgatg tccggcggtg
     3001 cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca
     3061 gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc
     3121 accttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     3181 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     3241 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     3301 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     3361 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     3421 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3481 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3541 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3601 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3661 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3721 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3781 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3841 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3901 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3961 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     4021 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     4081 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     4141 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4261 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4801 tcacatgtt
//
