LOCUS       sequence-238084-        5324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Entry Vector for Multisite Gateway Cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5324)
  AUTHORS   Gu Y, Innes RW
  TITLE     The KEEP ON GOING protein of Arabidopsis recruits the ENHANCED 
            DISEASE RESISTANCE1 protein to trans-Golgi network/early endosome 
            vesicles.
  JOURNAL   Plant Physiol. 2011 Apr;155(4):1827-38. doi: 10.1104/pp.110.171785. 
            Epub 2011 Feb 22.
  PUBMED    21343429
REFERENCE   2  (bases 1 to 5324)
  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"; doi: 
            "10.1104/pp.110.171785"; journalName: "Plant Physiol"; date: 
            "2011-04"; volume: "155"; issue: "4"; pages: "1827-38"
COMMENT     Sequence Label: pBSDONR P4r-P2
FEATURES             Location/Qualifiers
     source          1..5324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     238..258
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        240..258
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     295..311
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     295..311
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    361..592
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        737..839
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             840..1523
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(906..925)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1843..2148
                     /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     2027..2046
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    2496..2727
                     /gene="mutant version of attP"
                     /label=attP4
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2748..2767)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2749..2767)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2777..2794)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2777..2793)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2786..2808)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2934..3390)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3072..3093)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3284..3303
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3417..3521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3522..3590,3591..4382)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3740..3759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4553..5141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5042..5061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5295..5312
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      241 attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctctaga
      301 actagtggat cccccgggct gcaggaattc gatatcgcgg ccgccctgca gctggatggc
      361 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat
      421 gctttcttat aatgccaact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa
      481 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
      541 aacatatcca gtcactatga atcaactact tagatggtat tagtgacctg tagtcgacta
      601 agttggcagc atcacccgac gcactttgcg ccgaataaat acctgtgacg gaagatcact
      661 tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt
      721 ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
      781 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
      841 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
      901 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
      961 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1021 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     1081 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1141 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1201 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1261 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1321 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1381 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1441 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggg gggcgtaaac
     1501 gccgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     1561 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     1621 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     1681 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     1741 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     1801 aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac
     1861 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     1921 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     1981 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     2041 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
     2101 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     2161 cttatacaca gccagtctgc aggtcgatac agtagaaatt acagaaactt tatcacgttt
     2221 agtaagtata gaggctgaaa atccagatga agccgaacga cttgtaagag aaaagtataa
     2281 gagttgtgaa attgttcttg atgcagatga ttttcaggac tatgacacta gcgtatatga
     2341 ataggtagat gtttttattt tgtcacacaa aaaagaggct cgcacctctt tttcttattt
     2401 ctttttatga tttaatacgg cattgaggac aatagcgagt aggctggata cgacgattcc
     2461 gtttgagaag aacatttgga aggctgtcgg tcgacaaata atgattttat tttgactgat
     2521 agtgacctgt tcgttgcaac aaattgataa gcaatgcttt cttataatgc caactttgta
     2581 tagaaaagtt gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     2641 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatgaatcaa
     2701 ctacttagat ggtattagtg acctgtaggg gcccggtacc caattcgccc tatagtgagt
     2761 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2821 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2881 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt
     2941 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttta
     3001 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3061 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3121 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3181 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3241 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3301 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3361 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3421 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3481 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3541 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3601 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3661 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3721 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3781 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3841 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3901 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3961 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4021 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4081 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4141 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4201 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4261 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4321 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4381 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4441 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4501 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4561 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4621 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4681 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4741 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4801 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4861 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4921 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4981 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5041 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5101 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5161 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5221 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5281 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agag
//
