LOCUS       40924_37223        3813 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRL446, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3813)
  AUTHORS   Elhai J, Wolk CP.
  TITLE     A versatile class of positive-selection vectors based on the 
            nonviability of palindrome-containing plasmids that allows cloning 
            into long polylinkers
  JOURNAL   Gene 68 (1), 119-138 (1988)
  PUBMED    2851487
REFERENCE   2  (bases 1 to 3813)
  AUTHORS   Zhang W, Du Y, Khudyakov I, Fan Q, Gao H, Ning D, Wolk CP, Xu X.
  TITLE     A gene cluster that regulates both heterocyst differentiation and 
            pattern formation in Anabaena sp. strain PCC 7120
  JOURNAL   Mol. Microbiol. 66 (6), 1429-1443 (2007)
  PUBMED    18045384
REFERENCE   3  (bases 1 to 3813)
  AUTHORS   Elhai J, Xu X, Wolk CP.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2007) MSU-DOE Plant Research Laboratory and 
            Department of Plant Biology, Michigan State University, Wilson Road,
            E. Lansing, MI 48824, USA
REFERENCE   4  (bases 1 to 3813)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3813)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1988"; volume: "68"; issue: "1"; pages: "119-138"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Mol. 
            Microbiol."; date: "2007"; volume: "66"; issue: "6"; pages: 
            "1429-1443"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (15-DEC-2007) MSU-DOE Plant Research Laboratory and Department of 
            Plant Biology, Michigan State University, Wilson Road, E. Lansing, 
            MI 48824, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             562..1374
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1592..1608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1616..1632)
                     /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(1640..1670)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1685..1706)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1994..2582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2756..3613)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3614..3718)
                     /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 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaagggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagagt cgacctgcag gggggggggg gaaagccacg ttgtgtctca aaatctctga
      481 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
      541 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcgaggccg
      601 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
      661 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
      721 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      781 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
      841 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
      901 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
      961 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1021 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1081 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
     1141 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1201 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1261 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1321 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     1381 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcggc
     1441 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca
     1501 acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggact ctagaggatc
     1561 cccgggtacc gagctcgaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1681 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1741 gaaacctgtc gtgccagctg gattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1801 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1861 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1921 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2161 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     2221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2401 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2521 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2581 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2641 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2701 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2821 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2881 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2941 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3001 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3061 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3361 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3721 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3781 ataaaaatag gcgtatcacg aggccctttc gtc
//
