LOCUS       40924_14840        4962 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDM347, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4962)
  AUTHORS   Veltman DM, Akar G, Bosgraaf L, Van Haastert PJ.
  TITLE     A new set of small, extrachromosomal expression vectors for 
            Dictyostelium discoideum
  JOURNAL   Plasmid 61 (2), 110-118 (2009)
  PUBMED    19063918
REFERENCE   2  (bases 1 to 4962)
  AUTHORS   Veltman DM, Akar G, Bosgraaf L, van Haastert PJM.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUL-2008) Cell Biochemistry, Rijks Universiteit 
            Groningen, Kerklaan 30, Haren 9751 NN, The Netherlands
REFERENCE   3  (bases 1 to 4962)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4962)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2009"; volume: "61"; issue: "2"; pages: "110-118"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-JUL-2008) Cell Biochemistry, Rijks Universiteit Groningen, 
            Kerklaan 30, Haren 9751 NN, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..131
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        156..186
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             240..896
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1241..1543
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1587..1711)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2032..2048)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2522..2626
                     /label=AmpR promoter
     CDS             2627..3484
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3658..4246
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4534..4555
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4570..4600
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4608..4624
                     /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     4632..4648
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agatctacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
       61 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      121 cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt
      181 ataatgtgtg gattttgagt taggttccgt cgagattttc aggagctaag gaagctaaaa
      241 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
      301 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
      361 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
      421 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
      481 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
      541 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
      601 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
      661 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
      721 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
      781 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
      841 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa
      901 cgcgtggttc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt
      961 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat
     1021 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat
     1081 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt
     1141 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa
     1201 atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac
     1261 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     1321 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     1381 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     1441 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     1501 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct
     1561 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt
     1621 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     1681 tttctcgttc agctttcttg tacaaagtgg ttactagtac gcgtaaattg ttcacagaaa
     1741 atgtttctaa attatttaat aaataataaa aaaacaaatt gttgtaataa tctaatattt
     1801 tctttttttt ttaatttttt ttttttaaat cttaataatt attaagttat tttaattttt
     1861 tttttttttt tttttttttt tttttttttt tttctatcaa aaaaatcaaa tatatttaaa
     1921 aaatttatta tttacagata cattttgaat ggtgaagata aatatatgca ttagatgtaa
     1981 aacagccaaa gagtatgaaa atcaaaaaga tagccggcca tcaagcttgg cactggccgt
     2041 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2101 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2161 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2221 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2281 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2341 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2401 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2461 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2521 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2581 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2641 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2701 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2761 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2821 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2881 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2941 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3001 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3061 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3121 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3181 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3241 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3301 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3361 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3421 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3481 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3541 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3601 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3661 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3721 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3781 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3841 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3901 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3961 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4021 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4081 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4141 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4201 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4261 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4321 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4381 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     4441 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     4501 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4561 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4621 acaatttcac acaggaaaca gctatgacca tgattacgaa ttcgatggcc ggctaaaaaa
     4681 aatttttatt tatttttatt tattttgaat taaatagatt acaaattaat taatcccatc
     4741 aaatctttaa aaaaaaatgg tttaaaaaaa cttgggttgg ttaattatta tttgaaaatt
     4801 ttaaaaccca aattaaaaaa aaaaaatggg attcaaaaat tttttttttt tttttttttt
     4861 tttttttttt tttttttttt ttttcagatt gcataaaaag attttttttt tttttttttc
     4921 ttatttctta aaacaaataa attaaattaa aaaataaaaa tc
//
