LOCUS       40924_37183        4750 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRKW2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4750)
  AUTHORS   Wu W, Niehrs C.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2006) Division of Molecular Embryology (A050), 
            Deutsches Krebsforschungszetrum (DKFZ), Im Neuenheimer Feld 280, 
            Heidelberg 69120, Germany
REFERENCE   2  (bases 1 to 4750)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4750)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (26-MAY-2006) Division of Molecular Embryology (A050), Deutsches 
            Krebsforschungszetrum (DKFZ), Im Neuenheimer Feld 280, Heidelberg 
            69120, Germany"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(956..972)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1008..1142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1215..1572
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1592..1608)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1821..2276
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2608..2712
                     /label=AmpR promoter
     CDS             2713..3570
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3744..4332
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4620..4641
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4656..4686
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4694..4710
                     /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)."
     primer_bind     4718..4734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatccgggcc cgaatacaag ctacttgttc tttttgcact cgagggatcc
      961 actagttcta gagcggccgc caccgcggtg gagcttggcc gtggcccaac ttgtttattg
     1021 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1081 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     1141 tcgatcggga attaattcgg cgcagcacca tggcctgaaa taacctctga aagaggaact
     1201 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     1261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1321 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1381 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     1441 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     1501 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     1561 ggcttttgca aaaagctgtt aacagcttgg cactggccgt cgttttacaa cgtcgtgact
     1621 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatcccccc ttcgccagct
     1681 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgt agcctgaatg
     1741 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1801 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1861 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1921 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1981 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2041 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2101 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2161 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2221 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2281 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccaactcc gctatcgcta
     2341 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2401 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2461 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agtattcttg aagacgaaag
     2521 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2941 gcggtattat cccgtgatga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3241 gacaccacga tgccagcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     4141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4501 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4561 taatccagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4621 aatgtgagtt acctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     4681 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     4741 tacgaattaa
//
