LOCUS       62056_8430        9724 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 9724)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9724
                     /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             861..1649
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      2042..2294
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2640..2658)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2679..2695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2703..2719)
                     /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(2727..2758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2773..2794)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    3162..3186
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4486..5112
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5549..6613
                     /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      6682..6876
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7220..7360
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7546..8134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8224..9015)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9440..9464
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(9542..9716)
                     /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 attgaacaag atggattgca cgcaggttct ccggccgctt
      901 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
      961 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1021 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     1081 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     1141 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     1201 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     1261 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     1321 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     1381 aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga
     1441 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     1501 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     1561 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     1621 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     1681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     1741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     1801 catgctggag ttcttcgccc acgggatctc tgcggaacag gcggtcgaag gtgccgatat
     1861 cattacgaca gcaacggccg acaagcacaa cgccacgatc ctgagcgaca atatgatcgg
     1921 gcccggcgtc cacatcaacg gcgtcggcgg cgactgccca ggcaagaccg agatgcaccg
     1981 cgatatcttg ctgcgttcgg atattttcgt ggagttcccg ccacagaccc ggatgatccc
     2041 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     2101 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     2161 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     2221 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     2281 tatgttacta gatcacgcgt tttcgggttt acctcttcca gatacagctc aactgcaatg
     2341 cattaatgca ttggacctcg caaccctagt acgcccttca ggctccggcg aagcagaaga
     2401 atagcttagc agagtctatt ttcattttcg ggagacgaga tcaagcagat caacggtcgt
     2461 caagagacct acgagactga ggaatccgct cttggctcca cgcgactata tatttgtctc
     2521 taattgtact ttgacatgct cctcttcttt actctgatag cttgactatg aaaattccgt
     2581 caccagccct gactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     2641 ctttagtgag ggttaattcc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2701 aattgttatc cgctcacaat tccacacaac atacgagccg gaaghcataa agtgtaaagc
     2761 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2821 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2881 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3001 aggggataac gcaggaaaga acatgaaggc tggcgtaata gcgaagaggc ccgcaccgat
     3061 cgcccttccc aacagttgcg cagcctgaat ggcgaatgct agagcagctt gagcttggat
     3121 cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt ttgacaggat atattggcgg
     3181 gtaaacctaa gagaaaagag cgtttattag aataacggat atttaaaagg gcgtgaaaag
     3241 gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct cgggatcaaa
     3301 gtactttgat ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc
     3361 cgtcttctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc
     3421 ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact
     3481 agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc
     3541 gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct
     3601 gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca
     3661 ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg
     3721 cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc
     3781 tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga
     3841 ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg
     3901 ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg
     3961 cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg
     4021 ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag
     4081 tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg
     4141 acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga
     4201 cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg
     4261 gtacgtgttc gagccgcccg cgcacgtctc aaccgtgcgg ctgcatgaaa tcctggccgg
     4321 tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg
     4381 ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc
     4441 gtatatgatg cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct
     4501 gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc
     4561 ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc
     4621 gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg
     4681 attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc
     4741 caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg
     4801 cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc
     4861 attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc
     4921 acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag
     4981 tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt
     5041 gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa
     5101 tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa
     5161 gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca
     5221 cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga
     5281 tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc
     5341 taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc
     5401 ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga
     5461 acgggcggtt ggccaggcgt aagcggctgg gttgtctgcc ggccctgcaa tggcactgga
     5521 acccccaagc ccgaggaatc ggcgtgacgg tcgcaaacca tccggcccgg tacaaatcgg
     5581 cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg cccagcggca
     5641 acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg atcgaatccg
     5701 caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg
     5761 cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc gcgatagtcg
     5821 cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt
     5881 gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg ccggcatggc
     5941 cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg aatccatgaa
     6001 ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac acgttgcgga
     6061 cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc tggtagaaac
     6121 ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg
     6181 ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga tcgtaaagag
     6241 cgaaaccggg cggccggagt acatcgagat cgagctagct gattggatgt accgcgagat
     6301 cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt tgatcgatcc
     6361 cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg cagaagccag
     6421 atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca agaagttctg
     6481 tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga
     6541 ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg agggcgaagc
     6601 atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag caggggaaaa
     6661 aggtcgaaaa ggtctctttc ctgtggatag cacgtacatt gggaacccaa agccgtacat
     6721 tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc ggtcacacat
     6781 gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact ctttaaaact
     6841 tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc gcacagccga
     6901 agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg
     6961 tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag gcaatctacc
     7021 agggcgcgga caagccgcgc cgtcgccact cgaccgccgg cgcccacatc aaggcaccct
     7081 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     7141 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7201 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     7261 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7321 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     7381 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7441 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7501 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7561 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7621 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7681 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7741 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7801 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7861 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7921 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7981 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8041 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8101 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8161 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta
     8221 ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc
     8281 cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg
     8341 gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa
     8401 gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa
     8461 gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt
     8521 aaatggagtg tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa
     8581 gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc
     8641 gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg
     8701 ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct
     8761 ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca
     8821 tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt
     8881 catttttaaa tataggtttt cattttctcc caccagctta tataccttag caggagacat
     8941 tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat
     9001 tctcatttta gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa
     9061 gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa
     9121 taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg
     9181 gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa
     9241 catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc
     9301 cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc
     9361 cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg
     9421 ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac
     9481 aacttaataa cacattgcgg acgtttttaa tgtactgaat taacgccgaa ttaattcggg
     9541 ggatctggat tttagtactg gattttggtt ttaggaatta gaaattttat tgatagaagt
     9601 attttacaaa tacaaataca tactaagggt ttcttatatg ctcaacacat gagcgaaacc
     9661 ctataggaac cctaattccc ttatctggga actactcaca cattattatg gagaaacttg
     9721 gcaa
//
