LOCUS       62056_13600        9703 bp DNA     circular SYN 06-APR-2019
DEFINITION  Transformation vector pKW, complete sequence.
ACCESSION   MK424950
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9703)
  AUTHORS   Luo J, Shen P, Chen J.
  TITLE     A modular toolset of phiC31-based fluorescent protein tagging 
            vectors for Drosophila
  JOURNAL   Fly (Austin) (2019) In press
  PUBMED    30885036
REFERENCE   2  (bases 1 to 9703)
  AUTHORS   Luo J.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JAN-2019) Model Animal Research Center, Nanjing 
            University, 12 Xu-fu Rd Pu-kou Dist, Nanjing, Jiangsu 210061, China
REFERENCE   3  (bases 1 to 9703)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fly 
            (Austin) (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-JAN-2019) Model Animal Research Center, Nanjing University, 12 
            Xu-fu Rd Pu-kou Dist, Nanjing, Jiangsu 210061, China"
FEATURES             Location/Qualifiers
     source          1..9703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            666..4802
                     /label=mini-white
                     /note="This modified version of the white gene lacks part
                     of the first intron."
     protein_bind    4809..4842
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4843..5273
                     /label=gypsy insulator
                     /note="chromatin insulator from Drosophila"
     misc_feature    complement(5283..6663)
                     /label=K10 polyA signal sequence
                     /note="K10 polyA signal sequence"
     misc_feature    complement(6713..6757)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    6758..7188
                     /label=gypsy insulator
                     /note="gypsy insulator"
     protein_bind    7227..7296
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     promoter        complement(7515..7533)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7554..7570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7578..7594)
                     /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(7602..7632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7647..7668)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7956..8544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8718..9575)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9576..9680)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
      781 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
      841 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
      901 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
      961 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     1021 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     1081 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     1141 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     1201 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     1261 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     1321 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     1381 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     1441 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     1501 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     1561 cggggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     1621 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     1681 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     1741 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     1801 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     1861 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     1921 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     1981 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     2041 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     2101 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     2161 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccag gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggcactaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcatt taccgcccac agcgtcgtcc
     2461 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     2521 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     2581 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     2641 aagggtatct agcattacat tacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgctattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatttggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagtta gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaaaat aaataaattt catttaactc gcgaacatgt tgaagatatg
     3181 aatattaatg agatgcgagt aacattttaa tttgcagatg gttgccatct tgattggcct
     3241 catctttttg ggccaacaac tcacgcaagt gggcgtgatg aatatcaacg gagccatctt
     3301 cctcttcctg accaacatga cctttcaaaa cgtctttgcc acgataaatg taagtcttgt
     3361 ttagaataca tttgcatatt aataatttac taactttcta atgaatcgat tcgatttagg
     3421 tgttcacctc agagctgcca gtttttatga gggaggcccg aagtcgactt tatcgctgtg
     3481 acacatactt tctgggcaaa acgattgccg aattaccgct ttttctcaca gtgccactgg
     3541 tcttcacggc gattgcctat ccgatgatcg gactgcgggc cggagtgctg cacttcttca
     3601 actgcctggc gctggtcact ctggtggcca atgtgtcaac gtccttcgga tatctaatat
     3661 cctgcgccag ctcctcgacc tcgatggcgc tgtctgtggg tccgccggtt atcataccat
     3721 tcctgctctt tggcggcttc ttcttgaact cgggctcggt gccagtatac ctcaaatggt
     3781 tgtcgtacct ctcatggttc cgttacgcca acgagggtct gctgattaac caatgggcgg
     3841 acgtggagcc gggcgaaatt agctgcacat cgtcgaacac cacgtgcccc agttcgggca
     3901 aggtcatcct ggagacgctt aacttctccg ccgccgatct gccgctggac tacgtgggtc
     3961 tggccattct catcgtgagc ttccgggtgc tcgcatatct ggctctaaga cttcgggccc
     4021 gacgcaagga gtagccgaca tatatccgaa ataactgctt gttttttttt tttaccatta
     4081 ttaccatcgt gtttactgtt tattgccccc tcaaaaagct aatgtaatta tatttgtgcc
     4141 aataaaaaca agatatgacc tatagaatac aagtatttcc ccttcgaaca tccccacaag
     4201 tagactttgg atttgtcttc taaccaaaag acttacacac ctgcatacct tacatcaaaa
     4261 actcgtttat cgctacataa aacaccggga tatatttttt atatacatac ttttcaaatc
     4321 gcgcgccctc ttcataattc acctccacca caccacgttt cgtagttgct ctttcgctgt
     4381 ctcccacccg ctctccgcaa cacattcacc ttttgttcga cgaccttgga gcgactgtcg
     4441 ttagttccgc gcgattcggt tcgctcaaat ggttccgagt ggttcatttc gtctcaatag
     4501 aaattagtaa taaatatttg tatgtacaat ttatttgctc caatatattt gtatatattt
     4561 ccctcacagc tatatttatt ctaatttaat attatgactt tttaaggtaa ttttttgtga
     4621 cctgttcgga gtgattagcg ttacaatttg aactgaaagt gacatccagt gtttgttcct
     4681 tgtgtagatg catctcaaaa aaatggtggg cataatagtg ttgtttatat atatcaaaaa
     4741 taacaactat aataataaga atacatttaa tttagaaaat gcttggattt cactggaact
     4801 aggctagtat aacttcgtat aatgtatgct atacgaagtt attggccacg taataagtgt
     4861 gcgttgaatt tattcgcaaa aacattgcat attttcggca aagtaaaatt ttgttgcata
     4921 ccttatcaaa aaataagtgc tgcatacttt ttagagaaac caaataattt tttattgcat
     4981 acccgttttt aataaaatac attgcatacc ctcttttaat aaaaaatatt gcatactttg
     5041 acgaaacaaa ttttcgttgc atacccaata aaagattatt atattgcata cccgttttta
     5101 ataaaataca ttgcataccc tcttttaata aagaatattg catacgttga cgaaacaaat
     5161 tttcgttgca tacccaataa aagattatta tattgcatac cttttcttgc cataccattt
     5221 agccgatcaa ttgtgctcgg caacagtata tttgtggtgt gccaaccaac aacacgcgta
     5281 cttcagccaa tccgccgcac cctcagctcc aaagcacgca tatgacccgt gagcaggcag
     5341 agaaacgcag gcggatagga gtcactagcg cagttctgta tagcctgcac tatcaagccg
     5401 aagagctggt agatattgat gagcacatag tttctggtgg agtgcatcca gtcaacgccg
     5461 aaccaggccg gatacaggag ctctcgatcc gggcgaacga ccacgcggac gcaactcaaa
     5521 gtagtgccaa atacgaaaat actggcaaag gtgcggaaag tgccctgtgc gatattcact
     5581 tccttcctca aggcagaaat ctcctcggga tcgcccacca gccgggctct agcgtccatg
     5641 agccttagga gagagcgaat ctcatggagc tgcttcctca ccatgtacac attcgcaaac
     5701 ttcaaattgg ccacaatatc ggtaatagtt atggtcaggt tctcgcacaa tccggccatg
     5761 ttggtggtga acagcagcct ggccagcaag ctcaagggaa acagcacggt gaccaccaag
     5821 ttgaccgtaa tggagtatac cacgtaaagc aggcttgaaa cgcccggagg acgcatcagg
     5881 ccagtgagct cccaaacgcg ccagtggtag tacacagcac tgtgggtgtt cagtttgaaa
     5941 tcctcttgct tctccattgt ctcggttacc tttggtcaaa tccatgggtt ctattgccta
     6001 tatactcttg cgattaccag tgattgcgct attagctatt agatggattg ttggccaaac
     6061 ttgtcgctta agtggctggg aattgtaacc gtaggcccga gtgtaatgat cccccataaa
     6121 aagttttcgc aatgccttta ttttttgttg caaatctctc tttattctgc ggtattcttc
     6181 attattgcgg ggatggggaa agtgtttata tagaagcaac ttacgattga acccaaatgc
     6241 acctgacaag caaggtcaaa gggccagatt tttaaatata ttatttagtc ttaggactct
     6301 ctatttgcaa ttaaattact ttgctacctg agggttaaat cttccccatt gataataata
     6361 attccactat atgttcaatt gggtttcacc gcgcttagtt acatgacgag ccctaatgag
     6421 ccgtcggtgg tctataaact gtgccttaca aatacttgca actcttctcg ttttgaagtc
     6481 agcagagtta ttgctaattg ctaattgcta attgctttta actgatttct tcgaaattgg
     6541 tgctatgttt atggcgctat taacaagtat gaatgtcagg tttaaccagg ggatgcttaa
     6601 ttgtgttctc aacttcaaag gcagaaatgt ttactcttga ccatgggttt aggtataatg
     6661 ttatcaagct cctcgagtta acgttacgtt aacgttaacg ttcgaggtcg actctagact
     6721 cgagggtacc agatctgaat tcggatcctc aaggccttgg ccacgtaata agtgtgcgtt
     6781 gaatttattc gcaaaaacat tgcatatttt cggcaaagta aaattttgtt gcatacctta
     6841 tcaaaaaata agtgctgcat actttttaga gaaaccaaat aattttttat tgcatacccg
     6901 tttttaataa aatacattgc ataccctctt ttaataaaaa atattgcata ctttgacgaa
     6961 acaaattttc gttgcatacc caataaaaga ttattatatt gcatacccgt ttttaataaa
     7021 atacattgca taccctcttt taataaagaa tattgcatac gttgacgaaa caaattttcg
     7081 ttgcataccc aataaaagat tattatattg catacctttt cttgccatac catttagccg
     7141 atcaattgtg ctcggcaaca gtatatttgt ggtgtgccaa ccaacaacac tagagatcca
     7201 ctagtgtcga cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc
     7261 ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat gaacgggtcg
     7321 aggtggcggt agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc gccctcgaaa
     7381 ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc gggtgcggat
     7441 acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgac actagttcta
     7501 gccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     7561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7741 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     8401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8641 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8701 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8761 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8821 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8881 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8941 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     9001 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     9061 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     9121 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     9181 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9241 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9301 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9361 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9421 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9481 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9541 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9601 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9661 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
