LOCUS       62056_8420        9467 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9467)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..181
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        191..209
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     235..251
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        294..312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        654..837
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             858..1406
                     /codon_start=1
                     /label=BlpR
                     /note="phosphinothricin acetyltransferase"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      1785..2037
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2383..2401)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2422..2438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2446..2462)
                     /label=lac operator
                     /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(2470..2501)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2516..2537)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    2905..2929
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4229..4855
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5292..6356
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      6425..6619
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6963..7103
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7289..7877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7967..8758)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9183..9207
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(9285..9459)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 aacattaacg tttacaattt cgcgccattc gccattcagg ctgcgcaact gttgggaagg
       61 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      121 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      181 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
      241 cgacggtatc gataagcttg atatcgaatt cctgcagccc gggggatccg cgcaattaac
      301 cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgcga cgttaactga
      361 tattgaagga gcactttttg ggcttggctg gagctagtgg aggtcaacaa tgaatgccta
      421 ttttggttta gtcgtccagg cggtgagcac aaaatttgtg tcgtttgaca agatggttca
      481 tttaggcaac tggtcagatc agccccactt gtagcagtag cggcggcgct cgaagtgtga
      541 ctcttattag cagacaggaa ctaggacatt atcatcatct gctgcttggt gcacgataac
      601 ttggtgcgtt tgtcaagcaa ggtaagtgaa cgacccggtc ataccttctt aaggggtttc
      661 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
      721 taaggtcact atcagctagc aaatatttct tgtcaaaaat gctccactga cgttccataa
      781 attcccctcg gtatccaatt agagtctcat attcactctc aatccaaata atctgcaccg
      841 gatctggatc gtttcgcatg agcccagaac gacgcccggc cgacatccgc cgtgccaccg
      901 aggcggacat gccggcggtc tgcaccatcg tcaaccacta catcgagaca agcacggtca
      961 acttccgtac cgagccgcag gaaccgcagg agtggacgga cgacctcgtc cgtctgcggg
     1021 agcgctatcc ctggctcgtc gccgaggtgg acggcgaggt cgccggcatc gcctacgcgg
     1081 gcccctggaa ggcacgcaac gcctacgact ggacggccga gtcgaccgtg tacgtctccc
     1141 cccgccacca gcggacggga ctgggctcca cgctctacac ccacctgctg aagtccctgg
     1201 aggcacaggg cttcaagagc gtggtcgctg tcatcgggct gcccaacgac ccgagcgtgc
     1261 gcatgcacga ggcgctcgga tatgcccccc gcggcatgct gcgggcggcc ggcttcaagc
     1321 acgggaactg gcatgacgtg ggtttctggc agctggactt cagcctgccg gtaccgcccc
     1381 gtccggtcct gcccgtcacc gagatctgat tcgaaatgac cgaccaagcg acgcccaacc
     1441 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     1501 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     1561 cccacgggat ctctgcggaa caggcggtcg aaggtgccga tatcattacg acagcaacgg
     1621 ccgacaagca caacgccacg atcctgagcg acaatatgat cgggcccggc gtccacatca
     1681 acggcgtcgg cggcgactgc ccaggcaaga ccgagatgca ccgcgatatc ttgctgcgtt
     1741 cggatatttt cgtggagttc ccgccacaga cccggatgat ccccgatcgt tcaaacattt
     1801 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     1861 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     1921 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     1981 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcacg
     2041 cgttttcggg tttacctctt ccagatacag ctcaactgca atgcattaat gcattggacc
     2101 tcgcaaccct agtacgccct tcaggctccg gcgaagcaga agaatagctt agcagagtct
     2161 attttcattt tcgggagacg agatcaagca gatcaacggt cgtcaagaga cctacgagac
     2221 tgaggaatcc gctcttggct ccacgcgact atatatttgt ctctaattgt actttgacat
     2281 gctcctcttc tttactctga tagcttgact atgaaaattc cgtcaccagc cctgactagt
     2341 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     2401 tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2461 aattccacac aacatacgag ccggaaghca taaagtgtaa agcctggggt gcctaatgag
     2521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2761 agaacatgaa ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2821 gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg tcgtttcccg
     2881 ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     2941 gagcgtttat tagaataacg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     3001 tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc
     3061 ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac
     3121 atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt
     3181 cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac
     3241 gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac
     3301 caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc
     3361 gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta
     3421 cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac
     3481 ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag
     3541 ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt
     3601 gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag
     3661 gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc
     3721 cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg
     3781 catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc
     3841 aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc
     3901 gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt
     3961 ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc
     4021 ccgcgcacgt ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc
     4081 tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt
     4141 gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag
     4201 taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg
     4261 cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc
     4321 cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg
     4381 ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa
     4441 ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc
     4501 tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga
     4561 catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg
     4621 aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga
     4681 ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg
     4741 cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg
     4801 cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt
     4861 taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc
     4921 gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg
     4981 gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa
     5041 ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc
     5101 aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga
     5161 acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg
     5221 cgtaagcggc tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga
     5281 atcggcgtga cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg
     5341 acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag
     5401 cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac
     5461 cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt
     5521 ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg
     5581 ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc
     5641 cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg
     5701 acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga
     5761 agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc
     5821 ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca
     5881 ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat
     5941 ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg
     6001 agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc
     6061 cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc
     6121 tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga
     6181 tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc
     6241 tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc
     6301 cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat
     6361 gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct
     6421 ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt
     6481 acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa
     6541 aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa
     6601 cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc
     6661 ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg
     6721 ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg
     6781 cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt
     6841 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     6901 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     6961 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     7021 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     7081 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7921 acgaaaactc acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca
     7981 gtaaaatata atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata
     8041 gctcgacata ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt
     8101 cataccactt gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat
     8161 ctttcacaaa gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg
     8221 gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt
     8281 cccagttttc gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta
     8341 agcggctgtc taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc
     8401 tgatgcactc cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt
     8461 ccgagcaaag gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt
     8521 caaagtgcag gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt
     8581 cccgttccac atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt
     8641 tttcattttc tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta
     8701 cgcagcggta tttttcgatc agttttttca attccggtga tattctcatt ttagccattt
     8761 attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa
     8821 gacgaactcc aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt
     8881 ttcaaagttg ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc
     8941 gcggtgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga
     9001 gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac
     9061 atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg
     9121 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
     9181 ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacacattg
     9241 cggacgtttt taatgtactg aattaacgcc gaattaattc gggggatctg gattttagta
     9301 ctggattttg gttttaggaa ttagaaattt tattgataga agtattttac aaatacaaat
     9361 acatactaag ggtttcttat atgctcaaca catgagcgaa accctatagg aaccctaatt
     9421 cccttatctg ggaactactc acacattatt atggagaaac ttggcaa
//
