LOCUS       40924_44539        4039 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Low copy number bacterial vector for cloning sequences that cause 
            toxicity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4039)
  AUTHORS   TaKaRa
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4039)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(187..642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     855..871
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(875..931)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(941..957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(965..981)
                     /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(989..1019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1034..1055)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             1601..1789
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    1894..2034
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2220..2808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2982..3839)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3840..3944)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      241 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaatagc
      301 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      361 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      421 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      481 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      541 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      601 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
      661 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      721 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      781 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      841 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt gcatgcctgc aggtcgactc
      901 tagaggatcc ccgggtaccg agctcgaatt cgtaatcatg gtcatagctg tttcctgtgt
      961 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1021 cctggggtgc ctaatgagtg agctaactca cattacttgg cagaacatat ccatcgcgtc
     1081 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt
     1141 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     1201 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     1261 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     1321 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     1381 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     1441 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     1501 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     1561 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     1621 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     1681 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     1741 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     1801 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     1861 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     1921 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1981 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     2041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2641 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3841 ctcttctttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3961 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4021 atcacgaggc cctttcgtc
//
