LOCUS Exported 4773 bp ds-DNA circular SYN 10-AUG-2016 .
KEYWORDS pDONR-P2RP3
REFERENCE 1 (bases 1 to 4773)
AUTHORS Biofeng Lab.
FEATURES Location/Qualifiers
source 1..4773
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 268..295
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(591..822)
/gene="mutant version of attP"
/bound_moiety="BP Clonase(TM)"
/note="attP2"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
promoter 980..1082
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 1083..1742
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2084..2389
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2746..2977)
/gene="mutant version of attP"
/bound_moiety="BP Clonase(TM)"
/note="attP3"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(3015..3033)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3038..3054)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 3167..3976
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 4123..4711
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
601 taataccatc taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg
661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
721 ctcgttcaac tttcttgtac aaagttggca ttataaaaaa gcattgctta tcaatttgtt
781 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggagctct agagcgtcga
841 ctaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc
901 acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact
961 tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa
1021 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
1081 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
1141 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
1201 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
1261 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
1321 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
1381 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
1441 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
1501 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
1561 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
1621 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc
1681 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
1741 aatcgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
1801 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtgtgc
1861 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
1921 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
1981 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
2041 gaaatgaacg gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta
2101 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
2161 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
2221 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
2281 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
2341 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
2401 ccttatacac agccagtctg caggtcgata cagtagaaat tacagaaact ttatcacgtt
2461 tagtaagtat agaggctgaa aatccagatg aagccgaacg acttgtaaga gaaaagtata
2521 agagttgtga aattgttctt gatgcagatg attttcagga ctatgacact agcgtatatg
2581 aataggtaga tgtttttatt ttgtcacaca aaaaagaggc tcgcacctct ttttcttatt
2641 tctttttatg atttaatacg gcattgagga caatagcgag taggctggat acgacgattc
2701 cgtttgagaa gaacatttgg aaggctgtcg gtcgagctcg aattctacag gtcactaata
2761 ccatctaagt agttgattca tagtgactgc atatgttgtg ttttacagta ttatgtagtc
2821 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt
2881 tcaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat ttgttgcaac
2941 gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt agcgttaacg
3001 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
3061 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
3121 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
3181 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
3241 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
3301 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
3361 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
3421 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
3481 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
3541 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
3601 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
3661 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
3721 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
3781 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
3841 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
3901 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
3961 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
4021 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
4081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
4201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
4261 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
4321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
4381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
4441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
4501 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
4561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
4621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
4681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
4741 cctggccttt tgctggcctt ttgctcacat gtt
//
