LOCUS       62056_4620        9826 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 9826)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(65..239)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(325..670)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1200..1216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1224..1240)
                     /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(1248..1278)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1293..1314)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1505..2181
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     CDS             2250..3038
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3098..3272
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(3350..3374)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             3799..4590
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4680..5268
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5454..5594)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5938..6132)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6201..7265)
                     /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"
     CDS             complement(7702..8328)
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    complement(9629..9653)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca ccggtaattc
       61 gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt tattgataga
      121 agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa
      181 accctatagg aaccctaatt cccttatctg ggaactactc acacattatt atggagaaac
      241 tcgagcttgt cgatcgacaa gggactgacc acccggggat cctctagagt cccccgctgc
      301 aggttttcct gcaggagtcc cccgtgttct ctccaaatga aatgaacttc cttatataga
      361 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc
      421 acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct
      481 cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttcaa cgatggcctt
      541 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttccacta tcttcacaat
      601 aaagtgacag atagctgggc aatggaatcc gaggaggttt ccggatatta ccctttgttg
      661 aaaagtctca attgcccttt ggtcttctga gactgtatct ttgatatttt tggagtagac
      721 aagtgtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaagag
      781 actctgtatg aactgttcgc cagtctttac ggcgagttct gttaggtcct ctatttgaat
      841 ctttgactcc atggcctttg attcagtggg aactaccttt ttagagactc caatctctat
      901 tacttgcctt ggtttgtgaa gcaagccttg aatcgtccat actggaatag tacttctgat
      961 cttgagaaat atatctttct ctgtgttctt gatgcagtta gtcctgaatc ttttgactgc
     1021 atctttaacc ttcttgggaa ggtatttgat ttcctggaga ttattgctcg ggtagatcgt
     1081 cttgatgata cctgctgcgt aagcctctct aaccatctgt gggttagcat tctttctgaa
     1141 attgaaaagg caaatctggc gcgccgatcc ccgggtaccg agctcgaatt cgtaatcatg
     1201 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1261 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1321 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1381 ggccaacgcg cggggagagg cggtttgcgt attggctaga gcagcttgcc aacatggtgg
     1441 agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg
     1501 ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag
     1561 ctatctgtca cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc
     1621 attgcgataa aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg
     1681 gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
     1741 aagtggattg atgtgaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa
     1801 gatacagtct cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga
     1861 aacctcctcg gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag
     1921 gaaggtggca cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc
     1981 tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa
     2041 gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg
     2101 gatgacgcac aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt
     2161 catttggaga ggacacgctg aaatcaccag tctctctcta caaatctatc tctctcgagc
     2221 tttcgcagat ctgtcgatcg accatgggga ttgaacaaga tggattgcac gcaggttctc
     2281 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2341 ctgatgccgc cgtgtttcgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2401 acctgtccgg tgccctgaat gaactccagg acgaggcagc gcggctatcg tggctggcca
     2461 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2521 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2581 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2641 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2701 ttgtcgatca ggatgatctg gacgaagaac atcaggggct cgcgccagcc gaactgttcg
     2761 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacacat ggcgatgcct
     2821 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2881 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagaac
     2941 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3001 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgg
     3061 atcgatcctc tagctagagt cgatcgacaa gctcgagttt ctccataata atgtgtgagt
     3121 agttcccaga taagggaatt agggttccta tagggtttcg ctcatgtgtt gagcatataa
     3181 gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc
     3241 taaaaccaaa atccagtact aaaatccaga tcccccgaat taattcggcg ttaattcagt
     3301 acattaaaaa cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg tttacaccac
     3361 aatatatcct gccaccagcc agccaacagc tccccgaccg gcagctcggc acaaaatcac
     3421 cactcgatac aggcagccca tcagtccggg acggcgtcag cgggagagcc gttgtaaggc
     3481 ggcagacttt gctcatgtta ccgatgctat tcggaagaac ggcaactaag ctgccgggtt
     3541 tgaaacacgg atgatctcgc ggagggtagc atgttgattg taacgatgac agagcgttgc
     3601 tgcctgtgat caccgcggtt tcaaaatcgg ctccgtcgat actatgttat acgccaactt
     3661 tgaaaacaac tttgaaaaag ctgttttctg gtatttaagg ttttagaatg caaggaacag
     3721 tgaattggag ttcgtcttgt tataattagc ttcttggggt atctttaaat actgtagaaa
     3781 agaggaagga aataataaat ggctaaaatg agaatatcac cggaattgaa aaaactgatc
     3841 gaaaaatacc gctgcgtaaa agatacggaa ggaatgtctc ctgctaaggt atataagctg
     3901 gtgggagaaa atgaaaacct atatttaaaa atgacggaca gccggtataa agggaccacc
     3961 tatgatgtgg aacgggaaaa ggacatgatg ctatggctgg aaggaaagct gcctgttcca
     4021 aaggtcctgc actttgaacg gcatgatggc tggagcaatc tgctcatgag tgaggccgat
     4081 ggcgtccttt gctcggaaga gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg
     4141 tatgcggagt gcatcaggct ctttcactcc atcgacatat cggattgtcc ctatacgaat
     4201 agcttagaca gccgcttagc cgaattggat tacttactga ataacgatct ggccgatgtg
     4261 gattgcgaaa actgggaaga agacactcca tttaaagatc cgcgcgagct gtatgatttt
     4321 ttaaagacgg aaaagcccga agaggaactt gtcttttccc acggcgacct gggagacagc
     4381 aacatctttg tgaaagatgg caaagtaagt ggctttattg atcttgggag aagcggcagg
     4441 gcggacaagt ggtatgacat tgccttctgc gtccggtcga tcagggagga tatcggggaa
     4501 gaacagtatg tcgagctatt ttttgactta ctggggatca agcctgattg ggagaaaata
     4561 aaatattata ttttactgga tgaattgttt tagtacctag aatgcatgac caaaatccct
     4621 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4681 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4741 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4801 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4861 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4921 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4981 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5041 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5101 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5161 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5221 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5281 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5341 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5401 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gggcctgatg
     5461 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     5521 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     5581 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5641 tgctcccggc atccgcttac agacaagctg tgaccgtttc cgggagctgc atgtgtcaga
     5701 ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc cttgatgtgg gcgccggcgg
     5761 tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg taggccagcc
     5821 atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg tagggagcgc
     5881 agcgaccgaa gggtaggcgc tttttgcagc tcttgggctg tgcgctggcc agacagttat
     5941 gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta ggcggaaaaa
     6001 tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc aatgtacggc
     6061 tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt cccaatgtac
     6121 gtgctatcca caggaaagag gccttttcga cctttttccc ctgctagggc aatttgccct
     6181 agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag gttgcggtag
     6241 cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta ctccggcagg
     6301 tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc tccggcgctg
     6361 ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc tgcggcgcgg
     6421 cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta atcggggtga
     6481 accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca atcagctagc
     6541 tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta gcggctaatc
     6601 aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt cgtacgctgc
     6661 atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc tttctgcttt
     6721 ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg gaacacgcgg
     6781 ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag atgggaaacc
     6841 gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc tgcggaaacc
     6901 tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg gtcacgcttc
     6961 gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc cacgtcatag
     7021 agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct aatcgacggc
     7081 gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc cgcttgccac
     7141 gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg cgcggccttc
     7201 aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc ggatggtttg
     7261 cgaccgtcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcagac
     7321 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc
     7381 gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct
     7441 actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca gatagcagct
     7501 cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga tcctccgccg
     7561 gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc aacgttgcag
     7621 ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt ttgctcattt
     7681 tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca gcgcctggac
     7741 ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg cggcggcagt
     7801 gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct cgtacccggc
     7861 cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa
     7921 ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc
     7981 ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga agtcggctgc
     8041 cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc
     8101 gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt
     8161 tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg gatcggaatc
     8221 gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg ttgcgatggt
     8281 cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc gttccccttg
     8341 cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg caagctgttt
     8401 tactcaaata cacatcacct ttttagacgg cggcgctcgg tttcttcagc ggccaagctg
     8461 gccggccagg ccgccagctt ggcatcagac aaaccggcca ggatttcatg cagccgcacg
     8521 gttgagccgt gcgcgggcgg ctcgaacacg tacccggccg cgatcatctc cgcctcgatc
     8581 tcttcggtaa tgaaaaacgg ttcgtcctgg ccgtcctggt gcggtttcat gcttgttcct
     8641 cttggcgttc attctcggcg gccgccaggg cgtcggcctc ggtcaatgcg tcctcacgga
     8701 aggcaccgcg ccgcctggcc tcggtgggcg tcacttcctc gctgcgctca agtgcgcggt
     8761 acagggtcga gcgatgcacg ccaagcagtg cagccgcctc tttcacggtg cggccttcct
     8821 ggtcgatcag ctcgcgggcg tgcgcgatct gtgccggggt gagggtaggg cgggggccaa
     8881 acttcacgcc tcgggccttg gcggcctcgc gcccgctccg ggtgcggtcg atgattaggg
     8941 aacgctcgaa ctcggcaatg ccggcgaaca cggtcaacac catgcggccg gccggcgtgg
     9001 tggtgtcggc ccacggctct gccaggctac gcaggcccgc gccggcctcc tggatgcgct
     9061 cggcaatgtc cagtaggtcg cgggtgctgc gggccaggcg gtctagcctg gtcactgtca
     9121 caacgtcgcc agggcgtagg tggtcaagca tcctggccag ctccgggcgg tcgcgcctgg
     9181 tgccggtgat cttctcggaa aacagcttgg tgcagccggc cgcgtgcagt tcggcccgtt
     9241 ggttggtcaa gtcctggtcg tcggtgctga cgcgggcata gcccagcagg ccagcggcgg
     9301 cgctcttgtt catggcgtaa tgtctccggt tctagtcgca agtattctac tttatgcgac
     9361 taaaacacgc gacaagaaaa cgccaggaaa agggcagggc ggcagcctgt cgcgtaactt
     9421 aggacttgtg cgacatgtcg ttttcagaag acggctgcac tgaacgtcag aagccgactg
     9481 cactatagca gcggaggggt tggatcaaag tactttgatc ccgaggggaa ccctgtggtt
     9541 ggcatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgat
     9601 tattctaata aacgctcttt tctcttaggt ttacccgcca atatatcctg tcaaacactg
     9661 atagtttaaa ctgaaggcgg gaaacgacaa tctgatccaa gctcaagctg ctctagcatt
     9721 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     9781 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggt
//
