LOCUS       40924_48378        4789 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Donor vector pZErOTM7ZeoGFPattB DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  AUTHORS   Yonemura N, Tamura T, Uchino K, Kobayashi I, Tatematsu K, Iizuka T, 
            Sezutsu H, Muthulakshmi M, Nagaraju J, Kusakabe T.
  TITLE     PhiC31 integrase-mediated cassette exchange in silkworm embryos
  JOURNAL   Mol. Genet. Genomics 287 (9), 731-739 (2012)
  PUBMED    22842670
REFERENCE   2  (bases 1 to 4789)
  AUTHORS   Yonemura N, Kobayashi I, Uchino K, Sezutsu H, Tamura T, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-APR-2012) Contact:Naoyuki Yonemura National Institute 
            of Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba 
            Science City 305-8634, Japan
REFERENCE   3  (bases 1 to 4789)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4789)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Genet.
            Genomics"; date: "2012"; volume: "287"; issue: "9"; pages: "731-739"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-APR-2012) Contact:Naoyuki Yonemura National Institute of 
            Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba Science 
            City 305-8634, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_recomb     301..335
                     /label=Streptomyces phage phiC31 attB sequence
                     /note="Streptomyces phage phiC31 attB sequence"
     protein_bind    301..334
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth
                     et al., 2000)"
     polyA_signal    complement(439..560)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(624..1001)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MDAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWG
                     REFALRDPAGNCVHFVAEEQD"
     CDS             complement(624..995)
                     /codon_start=1
                     /gene="Zeo"
                     /product="Zeocin resistant protein"
                     /label=Zeo
                     /note="Zeocin region of GFP-Zeocin fusion gene"
                     /protein_id="BAM36531.1"
                     /translation="AKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDDV
                     TLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGRE
                     FALRDPAGNCVHFVAEEQD"
     gene            complement(624..995)
                     /gene="Zeo"
                     /label=Zeo
     CDS             complement(996..1700)
                     /codon_start=1
                     /label=Cycle 3 GFP
                     /note="Cycle 3 GFP (Crameri et al., 1996)"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMD"
     promoter        complement(1734..1781)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     protein_bind    complement(1823..1856)
                     /label=attB
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et
                     al., 2000)"
     promoter        complement(1881..1899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1906..1922)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2060..2359
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     rep_origin      2387..2766
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2817..3608)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      4020..4608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct aattctgcag atcccgatat
      301 cgcgcccggg gagcccaagg gcacgccctg gcacctcgaa attcgcttcg gtttatatga
      361 gacgagagta aggggtccgt caaaacaaaa catcgatgtt cccactggcc tggagcgact
      421 gtttttcagt agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      481 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      541 taagctgcaa taaacaagtt tcgaggtcga cccccctcgg acccgtcggg ccgcgtcgga
      601 ccggcggtgt tggtcggcgt cggtcagtcc tgctcctcgg ccacgaagtg cacgcagttg
      661 ccggccgggt cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg
      721 gccggcccgg aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc
      781 tcgtccaggc cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg
      841 accgcgctga tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg
      901 aagtcccggg agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc
      961 gcggtgagca ccggaacggc actggtcaac ttggcatcca tgccatgtgt aatcccagca
     1021 gcagttacaa actcaagaag gaccatgtgg tcacgctttt cgttgggatc tttcgaaagg
     1081 gcagattgtg tcgacaggta atggttgtct ggtaaaagga cagggccatc gccaattgga
     1141 gtattttgtt gataatggtc tgctagttga acggatccat cttcaatgtt gtgacgaatt
     1201 ttgaagttag ctttgattcc attcttttgt ttgtctgccg tgatgtatac attgtgtgag
     1261 ttatagttgt actcgagttt gtgtccgaga atgtttccat cttctttaaa atcaatacct
     1321 tttaactcga tacgattaac aagggtatca ccttcaaact tgacttcagc acgcgtcttg
     1381 tagttcccgt catctttgaa agatatagtg cgttcctgta cataaccttc gggcatggca
     1441 ctcttgaaaa agtcatgccg tttcatatga tccggataac gggaaaagca ttgaacacca
     1501 taagagaaag tagtgacaag tgttggccat ggaacaggta gttttccagt agtgcaaata
     1561 aatttaaggg taagctttcc gtatgtagca tcaccttcac cctctccact gacagaaaat
     1621 ttgtgcccat taacatcacc atctaattca acaagaattg ggacaactcc agtgaaaagt
     1681 tcttctcctt tgctagccat tttttctacc gccatggttt agttcctcac cttgtcgtat
     1741 tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctgc agcccggcag
     1801 atcccggcaa aggtgctgcg gccgcgcccg gggagcccaa gggcacgccc tggcaccgga
     1861 tcctagaggg cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     1921 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1981 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2041 gcgcagccta tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     2101 tctgtttgtg gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc
     2161 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     2221 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     2281 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     2341 cctgatgttc tggggaatat aaatgtcagg cctgaatggc gaatggacgc gccctgtagc
     2401 ggcgcattaa gcgcgcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2461 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2521 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagagct ttacggcacc
     2581 tcgaccgcaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2641 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2701 ctggaacaac actcaaccct atcgcggtct attcttttga tttataaggg atgttgccga
     2761 tttcggccta ttggttaaaa aatgagctga tttaacaaaa attttaacaa aattcagaag
     2821 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     2881 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     2941 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     3001 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     3061 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     3121 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     3181 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     3241 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     3301 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     3361 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     3421 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     3481 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     3541 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     3601 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat
     3661 cagatccttg gcggcgagaa agccatccag tttactttgc agggcttccc aaccttacca
     3721 gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc
     3781 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt
     3841 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     3901 ttctacgtga aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3961 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4021 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4081 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4141 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4201 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4261 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4321 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4381 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4441 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4501 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4561 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4621 gcggcctttt tacggttcct gggcttttgc tggccttttg ctcacatgtt ctttcctgcg
     4681 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4741 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
