LOCUS       40924_25551        4772 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pInSRT DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4772)
  AUTHORS   Shinomura T, Nakamura S, Ito K, Shirasawa S, Hook M, Kimura JH.
  TITLE     Adsorption of Follicular Dendritic Cell-secreted Protein (FDC-SP) 
            onto Mineral Deposits: APPLICATION OF A NEW STABLE GENE EXPRESSION 
            SYSTEM
  JOURNAL   J. Biol. Chem. 283 (48), 33658-33664 (2008)
  PUBMED    18806264
REFERENCE   2  (bases 1 to 4772)
  AUTHORS   Shinomura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JAN-2008) Contact:Tamayuki Shinomura Tokyo Medical and
            Dental University, Graduate School, Tissue Regeneration; 1-5-45 
            Yushima, Bunkyo-ku, Tokyo 113-8549, Japan
REFERENCE   3  (bases 1 to 4772)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4772)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2008"; volume: "283"; issue: "48"; pages: 
            "33658-33664"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JAN-2008) Contact:Tamayuki Shinomura Tokyo Medical and Dental 
            University, Graduate School, Tissue Regeneration; 1-5-45 Yushima, 
            Bunkyo-ku, Tokyo 113-8549, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..32
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    50..97
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    140..726
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             727..1323
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    1413..1637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(1671..1689)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1696..1712)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1853..2308)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2642..2746
                     /label=AmpR promoter
     CDS             2747..3604
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3778..4366
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4654..4675
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4690..4720
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4728..4744
                     /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     4752..4768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attacgccaa gctatttagg tgacactata gaatactcaa gcttcgaccg aagttcctat
       61 tccgaagttc ctattctcta gaaagtatag gaacttcgga tccccgggga gtcgacgtct
      121 cgaggaattc cgcccccccc ccctctccct cccccccccc taacgttact ggccgaagcc
      181 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
      241 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
      301 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
      361 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
      421 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
      481 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
      541 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
      601 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
      661 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
      721 acaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
      781 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
      841 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
      901 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
      961 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     1021 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     1081 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     1141 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     1201 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     1261 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     1321 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
     1381 tcgatgatct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1441 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1501 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1561 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1621 tgcggtgggc tctatggctt ctgaggcgga aagaacctgc agcaattcgc cctatagtga
     1681 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1741 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1801 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
     1861 aacgttaata ttttgttaaa attcgcgtta aatatttgtt aaatcagctc attttttaac
     1921 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     1981 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2041 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     2101 tttttgcggt cgaggtgccg taaagctcta aatcggaacc ctaaagggag cccccgattt
     2161 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2221 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2281 gcgcttaatg cgccgctaca gggcgcgtcg cctgatgcgg tattttctcc ttacgcatct
     2341 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2401 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2461 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2521 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2581 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2641 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2701 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2761 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2821 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2881 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2941 aatgatgagc acttttaaag ttctgctatg tcatacacta ttatcccgta ttgacgccgg
     3001 gcaagagcaa ctcggtcgcc gggcgcggta ttctcagaat gacttggttg agtactcacc
     3061 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3121 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3181 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3241 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatgcc
     3301 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3361 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3421 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3481 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3541 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3601 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3661 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3721 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3781 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3841 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3901 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3961 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4021 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4081 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4141 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4201 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4261 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4321 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tcgaaaaacg ccagcaacgc
     4381 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4441 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4501 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     4561 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     4621 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4681 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4741 acaatttcac acaggaaaca gctatgacca tg
//
