LOCUS       pBSDONR P1-P4                5371 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  ampicillin resistant derivative of pDONR221 P1-P4 for multisite 
            GATEWAY cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pBSDONR P1-P4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5371)
  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 5371)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..5371
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    669..900
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     CDS             complement(1296..1601)
                     /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     complement(1398..1417)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1951..2604)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     primer_bind     2519..2538
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2605..2707)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2852..3083)
                     /gene="mutant version of attP"
                     /label=attP4
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3133..3149)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3186..3206)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3186..3204)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3225..3241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3225..3241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3238..3260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3249..3265
                     /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)."
     promoter        complement(3273..3303)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3318..3339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3456..3473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3627..4215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3707..3726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4386..5246)
                     /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     5009..5028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5247..5351)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
      661 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
      721 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gctgaacgag aaacgtaaaa
      781 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
      841 taaaacacaa catatccagt cactatgaat caactactta gatggtatta gtgacctgta
      901 gtcgaccgac agccttccaa atgttcttcg ggtgatgctg ccaacttagt cgaccgacag
      961 ccttccaaat gttcttctca aacggaatcg tcgtatccag cctactcgct attgtcctca
     1021 atgccgtatt aaatcataaa aagaaataag aaaaagaggt gcgagcctct tttttgtgtg
     1081 acaaaataaa aacatctacc tattcatata cgctagtgtc atagtcctga aaatcatctg
     1141 catcaagaac aatttcacaa ctcttatact tttctcttac aagtcgttcg gcttcatctg
     1201 gattttcagc ctctatactt actaaacgtg ataaagtttc tgtaatttct actgtatcga
     1261 cctgcagact ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta
     1321 atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg
     1381 gagaccggca cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc
     1441 accaccgggt aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg
     1501 atcaccatcc gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga
     1561 taacggctct ctcttttata ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc
     1621 gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt
     1681 ttccgctttc cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac
     1741 cagaccggag atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact
     1801 gtcactgtaa tacgctgctt catagcatac ctctttttga catacttcgg gtatacatat
     1861 cagtatatat tcttataccg caaaaatcag cgcgcaaata cgcatactgt tatctggctt
     1921 ttagtaagcc ggatccacgc ggcgtttacg ccccccctgc cactcatcgc agtactgttg
     1981 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     2041 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     2101 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     2161 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     2221 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     2281 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     2341 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
     2401 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     2461 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     2521 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     2581 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     2641 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     2701 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     2761 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag
     2821 tgcgtcgggt gatgctgcca acttagtcga ctacaggtca ctaataccat ctaagtagtt
     2881 gattcatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2941 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcaa cttttctata
     3001 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     3061 cagtcaaaat aaaatcatta tttgccatcc agctgcaggg cggccgcgat atcgaattcc
     3121 tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag ctccagcttt
     3181 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     3241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5341 ggggttccgc gcacatttcc ccgaaaagtg c
//