LOCUS       62056_8450        9317 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 9317)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9317
                     /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        188..206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        246..264
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        309..659
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     CDS             664..1686
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(2233..2251)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2272..2288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2296..2312)
                     /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(2320..2351)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2366..2387)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    2755..2779
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4079..4705
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5142..6206
                     /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      6275..6469
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6813..6953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7139..7727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7817..8608)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9033..9057
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(9135..9309)
                     /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 tgaattgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgagggatc
      241 cgcgcaatta accctcacta aagggaacaa aagctggagc tccaccgcgg tggcggccgc
      301 gacgttaact gatattgaag gagcattttt gggcttggct ggagctagtg gaggtcaaca
      361 catcaatgct attttggttt agtcgtccag gcggatcaca aaatttgtgt cgtttgacaa
      421 gatggttcat ttaggcaact ggtcagatca gcccacttgt aagcagtagc ggcggcgctc
      481 gaagtgtgac tcttattagc agacaggaac gaggacatta ttatcatctg ctgcttggtg
      541 cacgataact tgtgcgtttg tcaagcaagg taagtgaacg acccggtcat accttcttaa
      601 gttcgccctt cctcccttta tttcagattc aatctgactt acctattcta cccaagcatc
      661 gatatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
      721 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
      781 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
      841 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
      901 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
      961 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     1021 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     1081 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     1141 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     1201 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     1261 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     1321 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     1381 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     1441 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     1501 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     1561 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     1621 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     1681 aaggaataga gtagatgccg accggatcga tccacttaac gttactgaaa tcatcaaaca
     1741 gcttgacgaa tctggatata agatcgttgg tgtcgatgtc agctccggag ttgagacaaa
     1801 tggtgttcag gatctcgata agatacgttc atttgtccaa gcagcaaaga gtgccttcta
     1861 gtgatttaat agctccatgt caacaagaat aaaacgcgtt ttcgggttta cctcttccag
     1921 atacagctca tctgcaatgc attaatgcat tgactgcaac ctagtaacgc cttcaggctc
     1981 cggcgaagag aagaatagct tagcagagct attttcattt tcgggagacg agatcaagca
     2041 gatcaacggt cgtcaagaga cctacgagac tgaggaatcc gctcttggct ccacgcgact
     2101 atatatttgt ctctaattgt actttgacat gctcctcttc tttactctga tagcttgact
     2161 atgaaaattc cgtcaccagc cctgactagt tctagagcgg ccgccaccgc ggtggagctc
     2221 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
     2281 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaaghca
     2341 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2401 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2581 tatccacaga atcaggggat aacgcaggaa agaacatgaa ggctggcgta atagcgaaga
     2641 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gctagagcag
     2701 cttgagcttg gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag
     2761 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataacg gatatttaaa
     2821 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     2881 cctcgggatc aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga
     2941 cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag
     3001 gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag
     3061 aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg
     3121 ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg
     3181 cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc
     3241 ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg
     3301 ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag
     3361 gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc
     3421 cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac
     3481 cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct
     3541 accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc
     3601 gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag
     3661 cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca
     3721 ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga
     3781 aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga
     3841 tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt ctcaaccgtg cggctgcatg
     3901 aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg
     3961 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc
     4021 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat
     4081 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca
     4141 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca
     4201 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat
     4261 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat
     4321 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt
     4381 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct
     4441 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg
     4501 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct
     4561 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg
     4621 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc
     4681 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc
     4741 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc
     4801 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac
     4861 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa
     4921 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc
     4981 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
     5041 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct gccggccctg
     5101 caatggcact ggaaccccca agcccgagga atcggcgtga cggtcgcaaa ccatccggcc
     5161 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     5221 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     5281 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
     5341 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     5401 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     5461 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     5521 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     5581 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     5641 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
     5701 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
     5761 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
     5821 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
     5881 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
     5941 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
     6001 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
     6061 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
     6121 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
     6181 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
     6241 tagcagggga aaaaggtcga aaaggtctct ttcctgtgga tagcacgtac attgggaacc
     6301 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
     6361 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
     6421 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
     6481 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
     6541 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
     6601 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
     6661 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6721 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6781 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     6841 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6901 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     6961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7561 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7801 tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
     7861 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
     7921 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
     7981 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
     8041 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
     8101 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
     8161 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
     8221 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
     8281 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
     8341 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
     8401 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
     8461 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
     8521 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
     8581 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
     8641 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
     8701 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
     8761 catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca
     8821 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
     8881 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
     8941 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
     9001 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
     9061 ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc
     9121 gaattaattc gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt
     9181 tattgataga agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca
     9241 catgagcgaa accctatagg aaccctaatt cccttatctg ggaactactc acacattatt
     9301 atggagaaac ttggcaa
//
