LOCUS       40924_17129        8865 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEGFP-2.attB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8865)
  AUTHORS   Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
  TITLE     A versatile platform for creating a comprehensive UAS-ORFeome 
            library in Drosophila
  JOURNAL   Development 140 (11), 2434-2442 (2013)
  PUBMED    23637332
REFERENCE   2  (bases 1 to 8865)
  AUTHORS   Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2013) Institute of Molecular Life Sciences, 
            University of Zurich, Winterthurerstrasse 190, Zurich 8057, 
            Switzerland
REFERENCE   3  (bases 1 to 8865)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8865)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development"; date: "2013"; volume: "140"; issue: "11"; pages: 
            "2434-2442"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-APR-2013) Institute of Molecular Life Sciences, University of 
            Zurich, Winterthurerstrasse 190, Zurich 8057, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..4136
                     /label=white dominant eye marker
                     /note="white dominant eye marker"
     CDS             join(479..550,952..1225,1300..1954,2016..2331,2552..2683,
                     2754..3368)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This ia a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4143..4176
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4189..4258
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             4264..4980
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     intron          5073..5138
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5268..5288
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    5560..5694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    5723..5792
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     promoter        complement(6011..6029)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6050..6066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6074..6090)
                     /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(6098..6128)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6143..6164)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6452..7040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7214..8071)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8072..8176)
                     /label=AmpR promoter
     rep_origin      complement(8202..8657)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8799..8815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8822..8840
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            8865
                     /label=mini-white
                     /note="This modified version of the white gene lacks part
                     of the first intron."
ORIGIN
        1 tagtatgtat gtaagttaat aaaacccatt tttgcggaaa gtagataaaa aaaacatttt
       61 ttttttttac tgcactggat atcattgaac ttatctgatc agttttaaat ttacttcgat
      121 ccaagggtat ttgatgtacc aggttctttc gattacctct cactcaaaat gacattccac
      181 tcaaagtcag cgctgtttgc ctccttctct gtccacagaa atatcgccgt ctctttcgcc
      241 gctgcgtccg ctatctcttt cgccaccgtt tgtagcgtta cgtagcgtca atgtccgcct
      301 tcagttgcat tttgtcagcg gtttcgtgac gaagctccaa gcggtttacg ccatcaatta
      361 aacacaaagt gctgtgccaa aactcctctc gcttcttatt tttgtttgtt ttttgagtga
      421 ttggggtggt gattggtttt gggtgggtaa gcaggggaaa gtgtgaaaaa tcccggcaat
      481 gggccaagag gatcaggagc tattaattcg cggaggcagc aaacacccat ctgccgagca
      541 tctgaacaat gtgagtagta catgtgcata catcttaagt tcacttgatc tataggaact
      601 gcgattgcaa catcaaattg tctgcggcgt gagaactgcg acccacaaaa atcccaaacc
      661 gcaattgcac aaacaaatag tgacacgaaa cagattattc tggtagctgt tctcgctata
      721 taagacaatt tttgagatca tatcatgatc aagacatcta aaggcattca ttttcgacta
      781 tattcttttt tacaaaaaat ataacaacca gatattttaa gctgatccta gatgcacaaa
      841 aaataaataa aagtataaac ctacttcgta ggatacttcg gggtactttt tgttcggggt
      901 tagatgagca taacgcttgt agttgatatt tgagatcccc tatcattgca gggtgacagc
      961 ggagcggctt cgcagagctg cattaaccag ggcttcgggc aggccaaaaa ctacggcacg
     1021 ctccggccac ccagtccgcc ggaggactcc ggttcaggga gcggccaact agccgagaac
     1081 ctcacctatg cctggcacaa tatggacatc tttggggcgg tcaatcagcc gggctccgga
     1141 tggcggcagc tggtcaaccg gacacgcgga ctattctgca acgagcgaca cataccggcg
     1201 cccaggaaac atttgctcaa gaacggtgag tttctattcg cagtcggctg atctgtgtga
     1261 aatcttaata aagggtccaa ttaccaattt gaaactcagt ttgcggcgtg gcctatccgg
     1321 gcgaactttt ggccgtgatg ggcagttccg gtgccggaaa gacgaccctg ctgaatgccc
     1381 ttgcctttcg atcgccgcag ggcatccaag tatcgccatc cgggatgcga ctgctcaatg
     1441 gccaacctgt ggacgccaag gagatgcagg ccaggtgcgc ctatgtccag caggatgacc
     1501 tctttatcgg ctccctaacg gccagggaac acctgatttt ccaggccatg gtgcggatgc
     1561 cacgacatct gacctatcgg cagcgagtgg cccgcgtgga tcaggtgatc caggagcttt
     1621 cgctcagcaa atgtcagcac acgatcatcg gtgtgcccgg cagggtgaaa ggtctgtccg
     1681 gcggagaaag gaagcgtctg gcattcgcct ccgaggcact aaccgatccg ccgcttctga
     1741 tctgcgatga gcccacctcc ggactggact catttaccgc ccacagcgtc gtccaggtgc
     1801 tgaagaagct gtcgcagaag ggcaagaccg tcatcctgac cattcatcag ccgtcttccg
     1861 agctgtttga gctctttgac aagatccttc tgatggccga gggcagggta gctttcttgg
     1921 gcactcccag cgaagccgtc gacttctttt cctagtgagt tcgatgtgtt tattaagggt
     1981 atctagcatt acattacatc tcaactccta tccagcgtgg gtgcccagtg tcctaccaac
     2041 tacaatccgg cggactttta cgtacaggtg ttggccgttg tgcccggacg ggagatcgag
     2101 tcccgtgatc ggatcgccaa gatatgcgac aattttgcta ttagcaaagt agcccgggat
     2161 atggagcagt tgttggccac caaaaatttg gagaagccac tggagcagcc ggagaatggg
     2221 tacacctaca aggccacctg gttcatgcag ttccgggcgg tcctgtggcg atcctggctg
     2281 tcggtgctca aggaaccact cctcgtaaaa gtgcgactta ttcagacaac ggtgagtggt
     2341 tccagtggaa acaaatgata taacgcttac aattcttgga aacaaattcg ctagatttta
     2401 gttagaattg cctgattcca cacccttctt agtttttttc aatgagatgt atagtttata
     2461 gttttgcaga aaataaataa atttcattta actcgcgaac atgttgaaga tatgaatatt
     2521 aatgagatgc gagtaacatt ttaatttgca gatggttgcc atcttgattg gcctcatctt
     2581 tttgggccaa caactcacgc aagtgggcgt gatgaatatc aacggagcca tcttcctctt
     2641 cctgaccaac atgacctttc aaaacgtctt tgccacgata aatgtaagtc ttgtttagaa
     2701 tacatttgca tattaataat ttactaactt tctaatgaat cgattcgatt taggtgttca
     2761 cctcagagct gccagttttt atgagggagg cccgaagtcg actttatcgc tgtgacacat
     2821 actttctggg caaaacgatt gccgaattac cgctttttct cacagtgcca ctggtcttca
     2881 cggcgattgc ctatccgatg atcggactgc gggccggagt gctgcacttc ttcaactgcc
     2941 tggcgctggt cactctggtg gccaatgtgt caacgtcctt cggatatcta atatcctgcg
     3001 ccagctcctc gacctcgatg gcgctgtctg tgggtccgcc ggttatcata ccattcctgc
     3061 tctttggcgg cttcttcttg aactcgggct cggtgccagt atacctcaaa tggttgtcgt
     3121 acctctcatg gttccgttac gccaacgagg gtctgctgat taaccaatgg gcggacgtgg
     3181 agccgggcga aattagctgc acatcgtcga acaccacgtg ccccagttcg ggcaaggtca
     3241 tcctggagac gcttaacttc tccgccgccg atctgccgct ggactacgtg ggtctggcca
     3301 ttctcatcgt gagcttccgg gtgctcgcat atctggctct aagacttcgg gcccgacgca
     3361 aggagtagcc gacatatatc cgaaataact gcttgttttt tttttttacc attattacca
     3421 tcgtgtttac tgtttattgc cccctcaaaa agctaatgta attatatttg tgccaataaa
     3481 aacaagatat gacctataga atacaagtat ttccccttcg aacatcccca caagtagact
     3541 ttggatttgt cttctaacca aaagacttac acacctgcat accttacatc aaaaactcgt
     3601 ttatcgctac ataaaacacc gggatatatt ttttatatac atacttttca aatcgcgcgc
     3661 cctcttcata attcacctcc accacaccac gtttcgtagt tgctctttcg ctgtctccca
     3721 cccgctctcc gcaacacatt caccttttgt tcgacgacct tggagcgact gtcgttagtt
     3781 ccgcgcgatt cggttcgctc aaatggttcc gagtggttca tttcgtctca atagaaatta
     3841 gtaataaata tttgtatgta caatttattt gctccaatat atttgtatat atttccctca
     3901 cagctatatt tattctaatt taatattatg actttttaag gtaatttttt gtgacctgtt
     3961 cggagtgatt agcgttacaa tttgaactga aagtgacatc cagtgtttgt tccttgtgta
     4021 gatgcatctc aaaaaaatgg tgggcataat agtgttgttt atatatatca aaaataacaa
     4081 ctataataat aagaatacat ttaatttaga aaatgcttgg atttcactgg aactaggcta
     4141 gcataacttc gtataatgta tgctatacga agttatgcta gcggatccaa gcttcgcagg
     4201 cctcgcaccg gtgaattcgt taacagatct gcggccgcgg ctcgagggta cctctagata
     4261 catatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4321 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4381 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4441 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4501 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4561 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4621 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4681 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4741 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4801 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4861 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4921 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4981 tgacctagga ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
     5041 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
     5101 gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg
     5161 aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc
     5221 catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca aaaaagaaga
     5281 gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg
     5341 tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa aaagctgcac
     5401 tgctatacaa gaaaattatg gaaaaatatt tgatgtatag tgccttgact agagatcata
     5461 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     5521 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     5581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccactag
     5701 tgtcgacgat gtaggtcacg gtctcgaagc cgcggtgcgg gtgccagggc gtgcccttgg
     5761 gctccccggg cgcgtactcc acctcaccca tctggtccat catgatgaac gggtcgaggt
     5821 ggcggtagtt gatcccggcg aacgcgcggc gcaccgggaa gccctcgccc tcgaaaccgc
     5881 tgggcgcggt ggtcacggtg agcacgggac gtgcgacggc gtcggcgggt gcggatacgc
     5941 ggggcagcgt cagcgggttc tcgacggtca cggcgggcat gtcgacacta gttctagcca
     6001 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     6061 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6121 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6181 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6241 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7141 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7201 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7261 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7321 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7381 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7441 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7501 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7561 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7621 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7681 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7741 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7801 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7861 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7921 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7981 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8041 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8101 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8161 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     8221 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     8281 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     8341 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     8401 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     8461 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     8521 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     8581 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     8641 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8701 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     8761 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     8821 gtaatacgac tcactatagg gcgaattggg tacgtaccgg gcccc
//
