LOCUS       V014895                 8089 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014895
VERSION     V014895
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8089)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8089
                     /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(591..1448)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1449..1553)
                     /label="AmpR promoter"
     rep_origin      1679..2267
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2453..2595)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2700..2888)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(3692..4306)
                     /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(5446..5523)
                     /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             6089..6799
                     /label="lambda repressor"
                     /note="phage lambda repressor"
     CDS             6800..6832
                     /label="ssrA tag (LVA)"
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness
                     et al., 2006)"
     terminator      6880..6951
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6967..6994
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     RBS             7070..7092
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7111..7128
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             7138..7155
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             7201..7908
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             7933..7950
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      8017..8064
                     /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 aattaattcg tatatatgag taaacttggt ctgacagtta ccaatgctta
      601 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      661 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      721 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      781 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      841 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      901 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      961 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1021 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1081 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1141 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1201 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1261 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1321 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1381 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1441 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1501 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacaaac
     1561 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     1621 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1681 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1741 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1801 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1861 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1921 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1981 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2041 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2101 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2161 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2221 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2281 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2341 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2401 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     2461 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag
     2521 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     2581 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2641 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2701 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     2761 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     2821 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     2881 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     2941 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3001 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3061 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3121 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     3181 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     3241 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     3301 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     3361 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     3421 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     3481 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     3541 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     3601 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     3661 taatgagtga gctaacttac atcaattgtc aatcgttgag catgccggcg cgcatcgcga
     3721 ggcgaaccag ctccgaaagg ctgttggcct gcatcttggt catgacgttg gcccgataca
     3781 cctcgatggt gcgcgggctg atgtcgtact cgcgggcgat cagcttgttg gaaaggccgg
     3841 cgatcagccc ttccatgacc tggcgctccc tggggctcaa cgaggcgacg cgggcggcga
     3901 tatcctgcgc gacggcctcg ctcttggcgg ccggctcggc ctggcggatc gccgattcga
     3961 tcatggcggt gaggcggtcg tcctcgaaag gcttttccag aaagtccacc gcccctaact
     4021 tcatcgcctc gaccgcgagc ggcacgtcgc cgtgaccggt catgatgagg atcggaaagg
     4081 ggctttgctg cgccttcatc cgcttcaaca gctcgatgcc gtcaaggccc ggcatgcgca
     4141 cgtcggagac gacgcagccg aaggagagac ccggcagggc gtcgagaaag gcttgcgcgt
     4201 cgtcaaacag cgtgacgccg aagccggcag aatccagcag gaaattcagc gaatcccgca
     4261 tcgccgcgtc gtcgtcgatg acgtagatat gtcccttggt cgtcatgcta gacctcctat
     4321 catctcgtcg gctgccggga gggtgaagcg gaaggtcgcc ccgcccgatg cgttgctctc
     4381 ggcccacatg cgcccgccgt gagcttcgat gatcgagcgg ctgatggaca gtcccacgcc
     4441 catgccggtg tccttggtgg tgaagaaagt ctgaaacagg ttcggaatga cgtcgtcctg
     4501 gaaaccgctg ccggtgtcgg acacttcgac ctcgatcatg tcgtcggcgg cgggggtgtt
     4561 ggtgacgacg agctcgcgtc gctgcgactg agccatcgct tccagcgcgt tgcggaacag
     4621 gttgaccagg acctgctgga tctgcacccg gtcggcgaga acgagatcgg cgcccggatc
     4681 gagactgaag cggagctgca cgttctgctc gcgcgcgccg gcaagcccga gcgcgccggc
     4741 ctcctcgatc agcttggaga gactctcgac ccgcttctcc gattcgccgc gggcaacgaa
     4801 gtcgcgcagg cgccggatga tctggccggc gcgcagcgcc tgctcggcgg cgcggtccag
     4861 ggcgctttcg accttcggtg tgttcggatc actgctgccg gcaagcagcc gccgcgagcc
     4921 cttcatgtag ttgctgatcg ccgccagcgg ctggttgagc tcgtgcgcga gcgcggacgc
     4981 catttcgccc atggcgctca gcctggagac gtggacgagc tcggattgca gttcctggag
     5041 gcgcgcctgg gtctgctggt gctcggtgat atcattctga ataccgacaa aatacgtttt
     5101 atcctctatt tccattggat caatatttaa ttcattccag aacatcgttc cgtctttttt
     5161 gtagttttgg atctgaacgg tgaccggttc tttattttgt aaagcggttc tgatgttgtc
     5221 cacttctgca ggatctgtgt gtttcccctg taagaagcga cagttctttc ctaaaatttc
     5281 ctcggtctcg tagccggtca tttgaacaaa gccttgattt acgtagacaa taggattatc
     5341 ttcaagtgcg ggatctgtaa ttaccacacc gactcgcacg tgatcaagtg cttttttgat
     5401 gacttccagc tgtcctggta tcccaaatga ttgaaaacta gccacattca ccaccctcaa
     5461 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     5521 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     5581 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     5641 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     5701 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     5761 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctacg
     5821 cccgtgatcc tgatcaccgg ctatccggac gaaaacatct cgacccgggc cgccgaggcc
     5881 ggcgtaaaag acgtggtttt gaagccgctt ctcgacgaaa acctgctcaa gcgtatccgc
     5941 cgcgccatcc aggaccggcc tcgggcatga cctacggggt tctacgtaag gcacccccct
     6001 taagatatcg ctcgaaattt tcgaacctcc cgataccgcg taccaatgcg tcatcacaac
     6061 ggagtctaga aaagaggaga aatactagat gagcacaaaa aagaaaccat taacacaaga
     6121 gcagcttgag gacgcacgtc gccttaaagc aatttatgaa aaaaagaaaa atgaacttgg
     6181 cttatcccag gaatctgtcg cagacaagat ggggatgggg cagtcaggcg ttggtgcttt
     6241 atttaatggc atcaatgcat taaatgctta taacgccgca ttgcttgcaa aaattctcaa
     6301 agttagcgtt gaagaattta gcccttcaat cgccagagaa atctacgaga tgtatgaagc
     6361 ggttagtatg cagccgtcac ttagaagtga gtatgagtac cctgtttttt ctcatgttca
     6421 ggcagggatg ttctcacctg agcttagaac ctttaccaaa ggtgatgcgg agagatgggt
     6481 aagcacaacc aaaaaagcca gtgattctgc attctggctt gaggttgaag gtaattccat
     6541 gaccgcacca acaggctcca agccgagctt tcctgacgga atgttaattc tcgttgaccc
     6601 tgagcaggct gttgagccag gtgatttctg catagccaga cttgggggtg atgagtttac
     6661 cttcaagaaa ctgatcaggg atagcggtca ggtgttttta caaccactaa acccacagta
     6721 cccaatgatc ccatgcaatg agagttgttc cgttgtgggg aaagttatcg ctagtcagtg
     6781 gcctgaagag acgtttggcg ctgcaaacga cgaaaactac gctttagtag cttaataacg
     6841 ctgatagtgc tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag
     6901 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     6961 cacactggct caccttcggg tgggcctttc tgcgtttata tactagagta acaccgtgcg
     7021 tgttgactat tttacctctg gcggtgataa tggttgctct agaaataatt ttgtttaact
     7081 ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg
     7141 gtgccgcgcg gcagccatat ggctagcatg actggtggac agcaaatggg tcggatccat
     7201 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     7261 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     7321 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     7381 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     7441 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     7501 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     7561 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     7621 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     7681 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     7741 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     7801 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     7861 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtg aaagcttgcg
     7921 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     7981 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     8041 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggat
//
