LOCUS       40924_48937        7480 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Retrofitting vector TKVBL-GVB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7480)
  AUTHORS   Kondo S, Booker M, Perrimon N.
  TITLE     Cross-species RNAi rescue platform in Drosophila melanogaster
  JOURNAL   Genetics 183 (3), 1165-1173 (2009)
  PUBMED    19720858
REFERENCE   2  (bases 1 to 7480)
  AUTHORS   Kondo S, Booker M, Perrimon N.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-FEB-2009) Department of Genetics, Harvard Medical 
            School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 7480)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7480)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics"; 
            date: "2009"; volume: "183"; issue: "3"; pages: "1165-1173"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-FEB-2009) Department of Genetics, Harvard Medical School, 77 
            Avenue Louis Pasteur, Boston, MA 02115, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1107..1140
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1457..1886
                     /label=gypsy insulator
                     /note="chromatin insulator from Drosophila"
     polyA_signal    complement(1894..2028)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2360..2755)
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(2772..3051)
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable
                     element copia (Sinclair et al., 1986)"
     polyA_signal    complement(3171..3252)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3381..4097)
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    4635..5019
                     /label=gypsy insulator
                     /note="chromatin insulator from Drosophila"
     CDS             complement(5391..6203)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6327..6962
                     /label=oriV
                     /note="incP origin of replication"
     protein_bind    6997..7066
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     mobile_element  complement(7292..7457)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ggtaccctgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct
       61 ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga
      121 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
      181 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
      241 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
      301 cagcgccctg caccattatg ttccggatct atgtcgggtg cggagaaaga ggtaatgaaa
      361 tggcagatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta aaagccttat
      421 atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt gctgatttat
      481 attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc ttagtacgtt
      541 agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat gagagcttag
      601 tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta ctatcaacag
      661 gttgaactgc tgatcttcgg atctatgtcg ggtgcggaga aagaggtaat gaaatggcag
      721 atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc
      781 ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat ttatattaat
      841 tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta cgttagccat
      901 gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc ttagtacgtt
      961 aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca acaggttgaa
     1021 ctgctgatct tcggatctat gtcgggtgcg gagaaagagg taatgaaatg gcatccggat
     1081 ctgcatcgca ggatgctgct ggctacataa cttcgtataa tgtatgctat acgaagttat
     1141 ctgtcatgcc atccgtatgt gggcggacaa taaagtctta aactgaacaa aatagatcta
     1201 aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt
     1261 atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga
     1321 agtgcaaatt gcccgtcgta ttaaagaggg gcgtggggtc gacgcggccg ctaactataa
     1381 cggtcctaag gtagcgagtt taaacgatcc tcgagttaac gttacgttaa cgttaacgtt
     1441 cgaggtcgac tctagttggc cacgtaataa gtgtgcgttg aatttattcg caaaaacatt
     1501 gcatattttc ggcaaagtaa aattttgttg cataccttat caaaaaataa gtgctgcata
     1561 ctttttagag aaaccaaata attttttatt gcatacccgt ttttaataaa atacattgca
     1621 taccctcttt taataaaaaa tattgcatac tttgacgaaa caaattttcg ttgcataccc
     1681 aataaaagat tattatattg catacccgtt tttaataaaa tacattgcat accctctttt
     1741 aataaaaata ttgcatacgt tgacgaaaca aattttcgtt gcatacccaa taaaagatta
     1801 ttatattgca taccttttct tgccatacca tttagccgat caattgtgct cggcaacagt
     1861 atatttgtgg tgtgccaacc aacaactcta gccgatccag acatgataag atacattgat
     1921 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1981 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttca agctagctgg
     2041 cctcgaggct gatctgcacc caatcggcag gcacgggcgg cgatctccaa tctgcgggat
     2101 cagtcagatc acccgagtgc gtgggcatga caatcgtgcc ctggggacca acacaatcca
     2161 gaagggcctg aatcactgcg accggccctc ccgcgaccca gccgagcgag cttagcgaac
     2221 tgtggacgag aactgtgcca ccaagcgtaa ggccgttctc tcgcatttgc cttgctaggc
     2281 tcgcgcgagt tgctggctga ggcgttctcg aaatcagctc ttgttcggtc ggcatctact
     2341 cccttgcgta taagggttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc
     2401 caactgccgt cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat
     2461 cgagaagaac ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga
     2521 tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga
     2581 gttctgcgca aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt
     2641 cgctagagag agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg
     2701 tagccgttgc tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatccctt
     2761 cgcataaagg gaacctgttg taatttataa tttatatttc ctttcttaat aaataaataa
     2821 atagtcaagt ttatgtttga gttttatgat ttatatttta agttatttca actgcaacac
     2881 cagcaccacg acctactcac agcaaaaaac gtacaagaag gaaagaagga ataaaaagag
     2941 tggtattctc ttacaatatg ttttatggca taaaaggtgt ggccattcat atcaaatata
     3001 aagtagtgtt gtttaacgtt atttttgtag gttgaatagt atattccaac agatgatgag
     3061 gggttcccaa tcctaaaccc atttgccgtt cccagaagca tgaaaccacc acgcaccgga
     3121 tcctagagga tccgatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3181 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3241 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3301 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcta
     3361 gctcgagttg cggccgctta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     3421 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     3481 gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     3541 ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg
     3601 aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgttg
     3661 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3721 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3781 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     3841 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg caggccgtag
     3901 cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgagc
     3961 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     4021 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     4081 tcgcccttgc tcaccatggt ggcggatccc ccgggctgca ggaattcgat atcaagctta
     4141 tcgataccgt cgaccggttt ggtgtctctg gattagacga ctgctggctg atggagcggc
     4201 tttgtgtcgg gaggagtatc cacacagcac aagaactcaa acggtagtga tatgaaaact
     4261 ggtcccgcag cccgttttat aaagcccgct tagcgctaaa accttatcag ccgcaatgct
     4321 ctccgctctc aagtcgcgtt caaaactttt accatatatg ggtatgaatt gttcgccgat
     4381 tgggttttca tttacggaga atttcctccg caactgggtg ttcgtgcttg tgtgtgtctt
     4441 gtatatcgcc atgaagagtg tacttccatg ggtcctatca gttgcggcat gtgattcaat
     4501 ttgggggtta ctcatattcg atacttcagc tggttatctt tatgctgtga tggtcacggt
     4561 gcttttacac attttttccc caactactca ttgtatgcca aaacatacat atggatccag
     4621 atccactagt tggccacgta ataagtgtgc gttgaattta ttcgcaaaaa cattgcatat
     4681 tttcggcaaa gtaaaatttt gttgcatacc ttatcaaaaa ataagtgctg catacttttt
     4741 agagaaacca aataattttt tattgcatac ccgtttttaa taaaatacat tgcataccct
     4801 cttttaataa aaaatattgc atactttgac gaaacaaatt ttcgttgcat acccaataaa
     4861 agattattat attgcatacc cgtttttaat aaaatacatt gcataccctc ttttaataaa
     4921 aatattgcat acgttgacga aacaaatttt cgttgcatac ccaataaaag attattatat
     4981 tgcatacctt ttcttgccat accatttagc cgatcaattg tgctcggcaa cagtatattt
     5041 gtggtgtgcc aaccaacaac tctagtgctg catccaacgc gttgggatcg gatccggccg
     5101 ccgctgaggt ctgcctcgtg aagaaggtgt tgctgactca taccaggcct gaatcgcccc
     5161 atcatccagc cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca
     5221 gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt
     5281 gatctgatcc ttcaactcag caagagttcg atttattcaa caaagccgcc gtcccgtcaa
     5341 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     5401 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     5461 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     5521 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     5581 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     5641 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     5701 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     5761 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     5821 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     5881 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     5941 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     6001 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     6061 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     6121 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac
     6181 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     6241 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     6301 acaacgtgct cgaaattccc ctttccgacg ctcaccgggc tggttgccct cgccgctggg
     6361 ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga
     6421 caccgcggcc ggccgccggc gttgtggata cctcgcggaa aacttggccc tcactgacag
     6481 atgaggggcg gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt tgacagatga
     6541 ggggcaggct cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa
     6601 cgcctgattt tacgcgagtt tcccacagat gatgtggaca agcctgggga taagtgccct
     6661 gcggtattga cacttgaggg gcgcgactac tgacagatga ggggcgcgat ccttgacact
     6721 tgaggggcag agtgctgaca gatgaggggc gcacctattg acatttgagg ggctgtccac
     6781 aggcagaaaa tccagcattt gcaagggttt ccgcccgttt ttcggccacc gctaacctgt
     6841 cttttaacct gcttttaaac caatatttat aaaccttgtt tttaaccagg gctgcgccct
     6901 gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc tcccggcccg
     6961 ctaacgcggg cctcggtcga cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca
     7021 gggcgtgccc ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat
     7081 gaacgggtcg aggtggcggt agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc
     7141 gccctcgaaa ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc
     7201 gggtgcggat acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgat
     7261 cacgatgaga atggccagac tagtgtcgac caaccagata agtgaaatct agttccaaac
     7321 tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt
     7381 ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac
     7441 tattttgtcc gcccacaccg taagatgctt ttctgtgact
//
