LOCUS       pAC10        8331 bp DNA     circular SYN 21-NOV-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8331)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 24, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
COMMENT     LOCUS       AF133829                8270 bp    DNA     circular SYN 
            20-MAR-2000
            DEFINITION  Shuttle vector pME6012, complete sequence.
            ACCESSION   AF133829
            VERSION     AF133829.1  GI:4512185
            KEYWORDS    .
            SOURCE      Shuttle vector pME6012
              ORGANISM  Shuttle vector pME6012
                        other sequences; artificial sequences; vectors.
            REFERENCE   1  (bases 1 to 8270)
              AUTHORS   Heeb,S., Itoh,Y., Nishijyo,T., Schnider,U., Keel,C., 
            Wade,J.,
                        Walsh,U., O'Gara,F. and Haas,D.
              TITLE     Small, stable shuttle vectors based on the minimal pVS1 
            replicon
                        for use in gram-negative, plant-associated bacteria
              JOURNAL   Mol. Plant Microbe Interact. 13 (2), 232-237 (2000)
               PUBMED   10659714
            REFERENCE   2  (bases 1 to 8270)
              AUTHORS   Heeb,S.
              TITLE     Direct Submission
              JOURNAL   Submitted (10-MAR-1999) Laboratoire de Biologie 
            Microbienne,
                        University of Lausanne, Batiment de Biologie, Lausanne 
            CH-1015,
                        Switzerland
FEATURES             Location/Qualifiers
     source          1..8331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             58..744
                     /codon_start=1
                     /label=ORF1;
                     /note="ORF1; not required for stable maintenance"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             741..956
                     /codon_start=1
                     /label=ORF2;
                     /note="ORF2; not required for stable maintenance"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             1043..1672
                     /codon_start=1
                     /label=StaA; similar to IncC2
                     /note="StaA; similar to IncC2"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1693..1908
                     /codon_start=1
                     /label=ORF3;
                     /note="ORF3; not required for stable maintenance"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    1939..1951
                     /label=KorB box
                     /note="KorB box"
     CDS             2101..3174
                     /codon_start=1
                     /label=high copy number RepA
                     /note="high copy number RepA"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     primer_bind     complement(2518..2537)
                     /label=AC_046
     primer_bind     2518..2537
                     /label=AC_047
     misc_difference 2837
                     /label=present
                     /note="present in pME290 and pME6010; results in decreased 
                     copy number"
     rep_origin      3240..3434
                     /direction=RIGHT
                     /label=pVS1 origin of replication
                     /note="pVS1 origin of replication"
     misc_feature    3240..3247
                     /label=DnaA box
                     /note="DnaA box"
     repeat_region   3251..3334
                     /label=21 bp repeats
                     /note="21 bp repeats"
     misc_feature    3335..3426
                     /label=AT-rich region
                     /note="AT-rich region"
     misc_feature    3427..3434
                     /label=DnaA box
                     /note="DnaA box"
     primer_bind     complement(4075..4097)
                     /label=AC_10
     primer_bind     4075..4097
                     /label=AC_018
     rep_origin      4127..4956
                     /direction=RIGHT
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     misc_feature    4931..4952
                     /label=p15A oriT
                     /note="p15A origin of transfer"
     misc_feature    5313..5365
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(6020..6039)
                     /label=AC_045
     misc_feature    6040..6137
                     /label=ConLS
     primer_bind     6040..6059
                     /label=AC_09
     primer_bind     complement(6072..6091)
                     /label=AC_019
     misc_feature    6145..6169
                     /label=ConLS scar
     misc_feature    6174..6243
                     /label=PA3
     promoter        6225
                     /label=E. coli promoter A3
     primer_bind     6244..6273
                     /label=AC_042
     RBS             6254..6261
     misc_feature    6270..6986
                     /label=E2-crimson
     primer_bind     6473..6492
                     /label=AC_024
     primer_bind     complement(6553..6572)
                     /label=AC_025
     terminator      6991..7073
                     /label=rpoC
     primer_bind     complement(7083..7143)
                     /label=AC_041
     CDS             complement(7144..8154)
                     /label=SpecR
     primer_bind     complement(7155..7174)
                     /label=AC_07
     promoter        complement(8155..8288)
                     /label=SpecR Promoter
     primer_bind     complement(8290..8312)
                     /label=AC_10
     primer_bind     8290..8312
                     /label=AC_018
     misc_feature    join(8313..8331,1..6039)
                     /label=pSV1
     primer_bind     join(8313..8331,1)
                     /label=AC_044
ORIGIN
        1 gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg
       61 aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac
      121 ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag
      181 atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct
      241 ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg
      301 gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg
      361 gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag
      421 ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga
      481 ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag
      541 ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc
      601 tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg
      661 cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat
      721 gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca
      781 ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc
      841 acgtctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg
      901 cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg
      961 tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata
     1021 aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc
     1081 aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt
     1141 tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga
     1201 tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat
     1261 cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc
     1321 cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg
     1381 ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct
     1441 acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc
     1501 cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag
     1561 ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc
     1621 tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga
     1681 ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc
     1741 agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac
     1801 tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag
     1861 accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga
     1921 ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc
     1981 aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag
     2041 cggctgggtt gtctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc
     2101 gtgagcggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg
     2161 gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc
     2221 cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg
     2281 gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc
     2341 gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt
     2401 ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac
     2461 gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg
     2521 gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga
     2581 gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga
     2641 gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg
     2701 cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag
     2761 ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac
     2821 atcgagatcg agctagttga ttggatgtac cgcgagatca cagaaggcaa gaacccggac
     2881 gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac
     2941 cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac
     3001 gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc
     3061 gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc
     3121 ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg
     3181 gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg tctctttcct
     3241 gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt
     3301 gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag
     3361 aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc
     3421 ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc
     3481 cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc
     3541 cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg
     3601 tcgccactcg accgccggcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     3661 cagggcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3721 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3781 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3841 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3901 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3961 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccttaa taagatgatc
     4141 ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt
     4201 gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct
     4261 tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt
     4321 caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc
     4381 tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg
     4441 ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc
     4501 gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga
     4561 acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4621 gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc
     4681 ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat
     4741 cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg
     4801 agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac
     4861 gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt
     4921 gccaacatag taagccagta tacactccgc tagcgctgag gtctgcctcg tgaagaaggt
     4981 gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca
     5041 cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc
     5101 acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt
     5161 cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat
     5221 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     5281 gttatgagcc atattcaacg ggaaacgtct tgctcgagat ctatcgatgc atgccatggt
     5341 acccgggagc tcgaattcga agcttttgcc attctcaccg gattcagtcg tcactcatgg
     5401 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     5461 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     5521 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     5581 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     5641 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata
     5701 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     5761 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
     5821 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca
     5881 gcaacacctt cttcacgagg cagacctcag cgccagaagg ccgccagaga ggccgagcgc
     5941 ggccgtgagg cttggacgct agggcagggc atgaaaaagc ccgtagcggg ctgctacggg
     6001 cgtctgacgc ggtggaaagg gggaggggat gttgtctaca tctgaatcat gcgcggatga
     6061 cacgaactca cgacggcgat cacagacatt aacccacagt acagacactg cgacaacgtg
     6121 gcaattcgtc gcaataccgt ctcactgaac tggccgataa ttgcagacga acgggtgaaa
     6181 caaaacggtt gacaacatga agtaaacacg gtacgatgta ccacatgaaa cgacagtgag
     6241 tcaatacaga aacagaggag atattacata tggatagcac tgagaacgtc atcaagccct
     6301 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6361 gcgtgggcga gggcaagccc tacgagggca cccagaccgc caagctgcaa gtgaccaagg
     6421 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcttctac ggctccaagg
     6481 cgtacatcaa gcaccccgcc gacatccccg actacctcaa gcagtccttc cccgagggct
     6541 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     6601 cctccctgca ggacggcacc ctcatctacc acgtgaagtt catcggcgtg aacttcccct
     6661 ccgacggccc cgtaatgcag aagaagactc tgggctggga gccctccact gagcgcaact
     6721 acccccgcga cggcgtgctg aagggcgaga accacatggc gctgaagctg aagggcggcg
     6781 gccactacct gtgtgagttc aagtccatct acatggccaa gaagcccgtg aagctgcccg
     6841 gctaccacta cgtggactac aagctcgaca tcacctccca caacgaggac tacaccgtgg
     6901 tggagcagta cgagcgcgcc gaggcccgcc accacctgtt ccagactcac ggtatggacg
     6961 aattgtacaa gcacgacgaa ttgtaaatcc gtaatcgtta atccgcaaat aacgtaaaaa
     7021 cccgcttcgg cgggtttttt tatgggggga gtttagggaa agagcatttg tcagctggat
     7081 cagcagatgg gaaccggata ctcctcagcc tccagagtag ccaatgagac gccgagcggc
     7141 cgcttatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     7201 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     7261 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     7321 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     7381 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     7441 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     7501 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     7561 ggctggctcg aagatacccg caagaatgtc attgcgctgc cattctccaa attgcagttc
     7621 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     7681 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     7741 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     7801 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     7861 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     7921 caccgcttcc ctcataatgt ttaactttgt tttagggcga ctgccctgct gcgtaacatc
     7981 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     8041 ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     8101 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     8161 cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg
     8221 tttccacggt gtgcgtcacc cggcaacctt gggtagcagc gaagtcgagg catttctgtc
     8281 ctggctggtc atgaccaaaa tcccttaacg tgttcctggc gttttcttgt c
//
