LOCUS       V013193                 7907 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013193
VERSION     V013193
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7907)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2271..2413)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(3510..4124)
                     /gene="fixJ"
                     /label="Transcriptional regulatory protein FixJ"
                     /note="Transcriptional regulatory protein FixJ from
                     Bradyrhizobium diazoefficiens (strain JCM 10833 / BCRC
                     13528 / IAM 13628 / NBRC 14792 / USDA 110). Accession#:
                     P23221"
     promoter        complement(5264..5341)
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5907..6617
                     /label="lambda repressor"
                     /note="phage lambda repressor"
     CDS             6618..6650
                     /label="ssrA tag (LVA)"
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness
                     et al., 2006)"
     terminator      6698..6769
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6785..6812
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     RBS             6888..6910
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6929..6946
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             6956..6973
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             7019..7726
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             7751..7768
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      7835..7882
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat caattgtcaa tcgttgagca tgccggcgcg catcgcgagg
     3541 cgaaccagct ccgaaaggct gttggcctgc atcttggtca tgacgttggc ccgatacacc
     3601 tcgatggtgc gcgggctgat gtcgtactcg cgggcgatca gcttgttgga aaggccggcg
     3661 atcagccctt ccatgacctg gcgctccctg gggctcaacg aggcgacgcg ggcggcgata
     3721 tcctgcgcga cggcctcgct cttggcggcc ggctcggcct ggcggatcgc cgattcgatc
     3781 atggcggtga ggcggtcgtc ctcgaaaggc ttttccagaa agtccaccgc ccctaacttc
     3841 atcgcctcga ccgcgagcgg cacgtcgccg tgaccggtca tgatgaggat cggaaagggg
     3901 ctttgctgcg ccttcatccg cttcaacagc tcgatgccgt caaggcccgg catgcgcacg
     3961 tcggagacga cgcagccgaa ggagagaccc ggcagggcgt cgagaaaggc ttgcgcgtcg
     4021 tcaaacagcg tgacgccgaa gccggcagaa tccagcagga aattcagcga atcccgcatc
     4081 gccgcgtcgt cgtcgatgac gtagatatgt cccttggtcg tcatgctaga cctcctatca
     4141 tctcgtcggc tgccgggagg gtgaagcgga aggtcgcccc gcccgatgcg ttgctctcgg
     4201 cccacatgcg cccgccgtga gcttcgatga tcgagcggct gatggacagt cccacgccca
     4261 tgccggtgtc cttggtggtg aagaaagtct gaaacaggtt cggaatgacg tcgtcctgga
     4321 aaccgctgcc ggtgtcggac acttcgacct cgatcatgtc gtcggcggcg ggggtgttgg
     4381 tgacgacgag ctcgcgtcgc tgcgactgag ccatcgcttc cagcgcgttg cggaacaggt
     4441 tgaccaggac ctgctggatc tgcacccggt cggcgagaac gagatcggcg cccggatcga
     4501 gactgaagcg gagctgcacg ttctgctcgc gcgcgccggc aagcccgagc gcgccggcct
     4561 cctcgatcag cttggagaga ctctcgaccc gcttctccga ttcgccgcgg gcaacgaagt
     4621 cgcgcaggcg ccggatgatc tggccggcgc gcagcgcctg ctcggcggcg cggtccaggg
     4681 cgctttcgac cttcggtgtg ttcggatcac tgctgccggc aagcagccgc cgcgagccct
     4741 tcatgtagtt gctgatcgcc gccagcggct ggttgagctc gtgcgcgagc gcggacgcca
     4801 tttcgcccat ggcgctcagc ctggagacgt ggacgagctc ggattgcagt tcctggaggc
     4861 gcgcctgggt ctgctggtgc tcggtgatat cattctgaat accgacaaaa tacgttttat
     4921 cctctatttc cattggatca atatttaatt cattccagaa catcgttccg tcttttttgt
     4981 agttttggat ctgaacggtg accggttctt tattttgtaa agcggttctg atgttgtcca
     5041 cttctgcagg atctgtgtgt ttcccctgta agaagcgaca gttctttcct aaaatttcct
     5101 cggtctcgta gccggtcatt tgaacaaagc cttgatttac gtagacaata ggattatctt
     5161 caagtgcggg atctgtaatt accacaccga ctcgcacgtg atcaagtgct tttttgatga
     5221 cttccagctg tcctggtatc ccaaatgatt gaaaactagc cacattcacc accctcaatt
     5281 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt
     5341 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     5401 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5461 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5521 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5581 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctacgcc
     5641 cgtgatcctg atcaccggct atccggacga aaacatctcg acccgggccg ccgaggccgg
     5701 cgtaaaagac gtggttttga agccgcttct cgacgaaaac ctgctcaagc gtatccgccg
     5761 cgccatccag gaccggcctc gggcatgacc tacggggttc tacgtaaggc acccccctta
     5821 agatatcgct cgaaattttc gaacctcccg ataccgcgta ccaatgcgtc atcacaacgg
     5881 agtctagaaa agaggagaaa tactagatga gcacaaaaaa gaaaccatta acacaagagc
     5941 agcttgagga cgcacgtcgc cttaaagcaa tttatgaaaa aaagaaaaat gaacttggct
     6001 tatcccagga atctgtcgca gacaagatgg ggatggggca gtcaggcgtt ggtgctttat
     6061 ttaatggcat caatgcatta aatgcttata acgccgcatt gcttgcaaaa attctcaaag
     6121 ttagcgttga agaatttagc ccttcaatcg ccagagaaat ctacgagatg tatgaagcgg
     6181 ttagtatgca gccgtcactt agaagtgagt atgagtaccc tgttttttct catgttcagg
     6241 cagggatgtt ctcacctgag cttagaacct ttaccaaagg tgatgcggag agatgggtaa
     6301 gcacaaccaa aaaagccagt gattctgcat tctggcttga ggttgaaggt aattccatga
     6361 ccgcaccaac aggctccaag ccgagctttc ctgacggaat gttaattctc gttgaccctg
     6421 agcaggctgt tgagccaggt gatttctgca tagccagact tgggggtgat gagtttacct
     6481 tcaagaaact gatcagggat agcggtcagg tgtttttaca accactaaac ccacagtacc
     6541 caatgatccc atgcaatgag agttgttccg ttgtggggaa agttatcgct agtcagtggc
     6601 ctgaagagac gtttggcgct gcaaacgacg aaaactacgc tttagtagct taataacgct
     6661 gatagtgcta gtgtagatcg ctactagagc caggcatcaa ataaaacgaa aggctcagtc
     6721 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     6781 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagtaac accgtgcgtg
     6841 ttgactattt tacctctggc ggtgataatg gttgctctag aaataatttt gtttaacttt
     6901 aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt
     6961 gccgcgcggc agccatatgg ctagcatgac tggtggacag caaatgggtc ggatccatat
     7021 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     7081 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     7141 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     7201 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     7261 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     7321 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     7381 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     7441 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     7501 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     7561 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     7621 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     7681 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtgaa agcttgcggc
     7741 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     7801 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     7861 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggat
//
