pDONR-P4P1R

LOCUS       Exported                4779 bp ds-DNA     circular SYN 10-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pDONR-P4P1R
  AUTHORS   Biofeng Lab.
FEATURES             Location/Qualifiers
     source          1..4779
                     /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    593..824
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP4"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1181..1486)
                     /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"
     CDS             complement(1836..2489)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(2490..2592)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    2750..2981
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP1"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     promoter        complement(3021..3039)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3044..3060)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3173..3982
                     /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      4129..4717
                     /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 tcgggcccga gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat
      721 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatg
      781 cagtcactat gaatcaacta cttagatggt attagtgacc tgtagaattc gagctcgacc
      841 gacagccttc caaatgttct tctcaaacgg aatcgtcgta tccagcctac tcgctattgt
      901 cctcaatgcc gtattaaatc ataaaaagaa ataagaaaaa gaggtgcgag cctctttttt
      961 gtgtgacaaa ataaaaacat ctacctattc atatacgcta gtgtcatagt cctgaaaatc
     1021 atctgcatca agaacaattt cacaactctt atacttttct cttacaagtc gttcggcttc
     1081 atctggattt tcagcctcta tacttactaa acgtgataaa gtttctgtaa tttctactgt
     1141 atcgacctgc agactggctg tgtataaggg agcctgacat ttatattccc cagaacatca
     1201 ggttaatggc gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga
     1261 taacggagac cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga
     1321 tatgcaccac cgggtaaagt tcacgggaga ctttatccga cagcagacgt gcactggcca
     1381 gggggatcac catccgtcgc ccgggcgtgt caataatatc actctgtaca tccacaaaca
     1441 gacgataacg gctctctctt ttataggtgt aaaccttaaa ctgcatttca ccagcccctg
     1501 ttctcgtcag caaaagagcc gttcatttca ataaaccggg cgacctcagc cgtcccttcc
     1561 tgattttccg ctttccagcg ttcggcacgc agacgacggg cttcattctg catggttgtg
     1621 cttaccagac cggagatatt gacatcatat atgccttgag caactgatag ctgtcgctgt
     1681 caactgtcac tgtaatacgc tgcttcatag catacctctt tttgacatac ttcgggtata
     1741 catatcagta tatattctta taccgcaaaa atcagcgcgc aaatacgcat actgttatct
     1801 ggcttttagt aagccggatc cacgcggcgt ttacgccccc cctgccactc atcgcagtac
     1861 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     1921 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     1981 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     2041 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     2101 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     2161 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     2221 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     2281 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     2341 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     2401 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     2461 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     2521 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     2581 acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca
     2641 gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg
     2701 caaagtgcgt cgggtgatgc tgccaactta gtcgacgctc tagagctcca aataatgatt
     2761 ttattttgac tgatagtgac ctgttcgttg caacaaattg ataagcaatg cttttttata
     2821 atgccaactt tgtacaaaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     2881 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatgcag
     2941 tcactatgaa ccaactactt agatggtatt agtgacctgt agaattcgag ctctagagct
     3001 gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     3061 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3121 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3181 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3241 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3301 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3361 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3421 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3481 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3541 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3601 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3661 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3721 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3781 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3841 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3901 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3961 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     4021 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     4081 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4261 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4501 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4741 acggttcctg gccttttgct ggccttttgc tcacatgtt
//