LOCUS       Exported                6693 bp ds-DNA     circular SYN 01-AUG-2024
DEFINITION  Gateway Expression Clone,.
ACCESSION   V1050
VERSION     .
KEYWORDS    pGL4_10xUAS-CMVmin-BC0431-luc2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     MASTER for EXT design
FEATURES             Location/Qualifiers
     source          1..6693
                     /organism="synthetic DNA construct"
     protein_bind    47..71
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        101..203
                     /gene="enhancer"
                     /label=5xUAS
                     /note="5xUAS"
                     /note="/SECDrawas=Region"
     misc_feature    101..117
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    120..136
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    144..160
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    163..179
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    187..203
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     enhancer        235..337
                     /gene="enhancer"
                     /label=5xUAS
                     /note="5xUAS"
                     /note="/SECDrawas=Region"
     misc_feature    235..251
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    254..270
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    278..294
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    297..313
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     misc_feature    321..337
                     /label=1xUAS segment, CGGRNNRCYNYNCNCCG
                     /note="1xUAS segment, CGGRNNRCYNYNCNCCG"
     protein_bind    complement(358..378)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        394..431
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    514..534
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    535..687
                     /gene="extassay structure"
                     /label=AFA EXT cassette
                     /note="AFA EXT cassette"
                     /note="/SECDrawas=Region"
     misc_feature    535..560
                     /gene="extassay structure"
                     /label=Decoding primer fwd
                     /note="Decoding primer fwd"
                     /note="/SECDrawas=Region"
     promoter        561..579
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        581..593
                     /gene="promoter"
                     
                     /note="truncated SP6"
                     /note="/SECDrawas=Region"
     misc_feature    594..642
                     /label=E00431
     promoter        complement(643..661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(662..687)
                     /gene="extassay structure"
                     /label=Decoding Primer rev
                     /note="Decoding Primer rev"
                     /note="/SECDrawas=Region"
     protein_bind    complement(685..709)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      767..776
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             773..2425
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     1034..1051
                     /label=luc2_pGL4_1 forw
     primer_bind     1115..1132
                     /label=luc2_pGL4_1 rev
     primer_bind     1336..1353
                     /label=luc2_pGL4_2 forw
     primer_bind     1634..1651
                     /label=luc2_pGL4_3 forw
     primer_bind     1947..1965
                     /label=luc2_pGL4_4 forw
     primer_bind     2234..2251
                     /label=luc2_pGL4_5 forw
     misc_feature    2426..2602
                     /gene="miscellaneous"
                     /label=not translated CP destablisation signal
                     /note="not translated CP destablisation signal"
                     /note="/SECDrawas=Region"
     CDS             2429..2476
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             2480..2599
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2639..2860
                     /gene="polya tail"
                     /label=SV40 late
                     /note="SV40 late"
                     /note="/SECDrawas=Region"
     polyA_signal    2648..2769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2908..3326
                     /gene="promoter"
                     /note="SV40"
                     /note="/SECDrawas=Region"
     promoter        2963..3320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3171..3306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3351..4388
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4367..4375
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4412..4460
                     /note="synthetic polyadenylation signal"
     primer_bind     4527..4546
                     /label=Reporter Vector primer 4
                     /note="Reporter Vector primer 4 (RVprimer4) binding region"
     rep_origin      complement(4772..5391)
                     /direction=LEFT
                     /gene="origin of replication"
                     /label=pBR322
                     /note="pBR322"
                     /note="/SECDrawas=Region"
     rep_origin      complement(4787..5375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5575..6435)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    6540..6693
                     /gene="polya tail"
                     /label=transcriptional pause site
                     /note="transcriptional pause site"
                     /note="/SECDrawas=Region"
     polyA_signal    6540..6588
                     /note="synthetic polyadenylation signal"
     misc_feature    6602..6693
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6642..6661
                     /label=Reporter Vector primer 3
                     /note="Reporter Vector primer 3 (RVprimer3) binding region"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaaacaa gtttgtacaa
       61 aaaagcaggc tccctaactg gccggtaccg agtttctaga cggagtactg tcctccgagc
      121 ggagtactgt cctccgactc gagcggagta ctgtcctccg atcggagtac tgtcctccgc
      181 gaattccgga gtactgtcct ccgaagacgc tagtgccggt accgagtttc tagacggagt
      241 actgtcctcc gagcggagta ctgtcctccg actcgagcgg agtactgtcc tccgatcgga
      301 gtactgtcct ccgcgaattc cggagtactg tcctccgaag acgctagcgg ggggctacaa
      361 cttttctata caaagttgcg gtacccgggt cgagtaggcg tgtacggtgg gaggcctata
      421 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
      481 acctccatag aagacaccgg gaccgatcca gcccaacttt gtataataaa gttgagctag
      541 ttgctaagtc tgccgagtag aattaaccct cactaaaggg taggtgacac tattctatac
      601 tatcaacatc tttagaccgt gtcttttcta tactaatctt accctatagt gagtcgtatt
      661 agtagacgcg agtcaatgca tgtacgacag ctttcttgta caaagtggtt cgatatcaag
      721 atctggcctc ggcggccaag cttggcaatc cggtactgtt ggtaaagcca ccatggaaga
      781 tgccaaaaac attaagaagg gcccagcgcc attctaccca ctcgaagacg ggaccgccgg
      841 cgagcagctg cacaaagcca tgaagcgcta cgccctggtg cccggcacca tcgcctttac
      901 cgacgcacat atcgaggtgg acattaccta cgccgagtac ttcgagatga gcgttcggct
      961 ggcagaagct atgaagcgct atgggctgaa tacaaaccat cggatcgtgg tgtgcagcga
     1021 gaatagcttg cagttcttca tgcccgtgtt gggtgccctg ttcatcggtg tggctgtggc
     1081 cccagctaac gacatctaca acgagcgcga gctgctgaac agcatgggca tcagccagcc
     1141 caccgtcgta ttcgtgagca agaaagggct gcaaaagatc ctcaacgtgc aaaagaagct
     1201 accgatcata caaaagatca tcatcatgga tagcaagacc gactaccagg gcttccaaag
     1261 catgtacacc ttcgtgactt cccatttgcc acccggcttc aacgagtacg acttcgtgcc
     1321 cgagagcttc gaccgggaca aaaccatcgc cctgatcatg aacagtagtg gcagtaccgg
     1381 attgcccaag ggcgtagccc taccgcaccg caccgcttgt gtccgattca gtcatgcccg
     1441 cgaccccatc ttcggcaacc agatcatccc cgacaccgct atcctcagcg tggtgccatt
     1501 tcaccacggc ttcggcatgt tcaccacgct gggctacttg atctgcggct ttcgggtcgt
     1561 gctcatgtac cgcttcgagg aggagctatt cttgcgcagc ttgcaagact ataagattca
     1621 atctgccctg ctggtgccca cactatttag cttcttcgct aagagcactc tcatcgacaa
     1681 gtacgaccta agcaacttgc acgagatcgc cagcggcggg gcgccgctca gcaaggaggt
     1741 aggtgaggcc gtggccaaac gcttccacct accaggcatc cgccagggct acggcctgac
     1801 agaaacaacc agcgccattc tgatcacccc cgaaggggac gacaagcctg gcgcagtagg
     1861 caaggtggtg cccttcttcg aggctaaggt ggtggacttg gacaccggta agacactggg
     1921 tgtgaaccag cgcggcgagc tgtgcgtccg tggccccatg atcatgagcg gctacgttaa
     1981 caaccccgag gctacaaacg ctctcatcga caaggacggc tggctgcaca gcggcgacat
     2041 cgcctactgg gacgaggacg agcacttctt catcgtggac cggctgaaga gcctgatcaa
     2101 atacaagggc taccaggtag ccccagccga actggagagc atcctgctgc aacaccccaa
     2161 catcttcgac gccggggtcg ccggcctgcc cgacgacgat gccggcgagc tgcccgccgc
     2221 agtcgtcgtg ctggaacacg gtaaaaccat gaccgagaag gagatcgtgg actatgtggc
     2281 cagccaggtt acaaccgcca agaagctgcg cggtggtgtt gtgttcgtgg acgaggtgcc
     2341 taaaggactg accggcaagt tggacgcccg caagatccgc gagattctca ttaaggccaa
     2401 gaagggcggc aagatcgccg tgtaatctgc ttgcaagaac tggttcagta gcttaagcca
     2461 ctttgtgatc caccttaaca gccacggctt ccctcccgag gtggaggagc aggccgccgg
     2521 caccctgccc atgagctgcg cccaggagag cggcatggat agacaccctg ctgcttgcgc
     2581 cagcgccagg atcaacgtct aactgcagtc tagagtcggg gcggccggcc gcttcgagca
     2641 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2701 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2761 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2821 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgttt
     2881 gcgtattggg cgctcttccg ctgatctgcg cagcaccatg gcctgaaata acctctgaaa
     2941 gaggaacttg gttagctacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3001 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3181 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3241 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3301 gaggcctagg cttttgcaaa aagctcgatt cttctgacac tagcgccacc atgaagaagc
     3361 ccgaactcac cgctaccagc gttgaaaaat ttctcatcga gaagttcgac agtgtgagcg
     3421 acctgatgca gttgtcggag ggcgaagaga gccgagcctt cagcttcgat gtcggcggac
     3481 gcggctatgt actgcgggtg aatagctgcg ctgatggctt ctacaaagac cgctacgtgt
     3541 accgccactt cgccagcgct gcactaccca tccccgaagt gttggacatc ggcgagttca
     3601 gcgagagcct gacatactgc atcagtagac gcgcccaagg cgttactctc caagacctcc
     3661 ccgaaacaga gctgcctgct gtgttacagc ctgtcgccga agctatggat gctattgccg
     3721 ccgccgacct cagtcaaacc agcggcttcg gcccattcgg gccccaaggc atcggccagt
     3781 acacaacctg gcgggatttc atttgcgcca ttgctgatcc ccatgtctac cactggcaga
     3841 ccgtgatgga cgacaccgtg tccgccagcg tagctcaagc cctggacgaa ctgatgctgt
     3901 gggccgaaga ctgtcccgag gtgcgccacc tcgtccatgc cgacttcggc agcaacaacg
     3961 tcctgaccga caacggccgc atcaccgccg taatcgactg gtccgaagct atgttcgggg
     4021 acagtcagta cgaggtggcc aacatcttct tctggcggcc ctggctggct tgcatggagc
     4081 agcagactcg ctacttcgag cgccggcatc ccgagctggc cggcagccct cgtctgcgag
     4141 cctacatgct gcgcatcggc ctggatcagc tctaccagag cctcgtggac ggcaacttcg
     4201 acgatgctgc ctgggctcaa ggccgctgcg atgccatcgt ccgcagcggg gccggcaccg
     4261 tcggtcgcac acaaatcgct cgccggagcg cagccgtatg gaccgacggc tgcgtcgagg
     4321 tgctggccga cagcggcaac cgccggccca gtacacgacc gcgcgctaag gaggtaggtc
     4381 gagtttaaac tctagaaccg gtcatggccg caataaaata tctttatttt cattacatct
     4441 gtgtgttggt tttttgtgtg ttcgaactag atgctgtcga ccgatgccct tgagagcctt
     4501 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     4561 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     4621 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4681 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4741 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4801 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4861 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4921 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4981 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5041 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5101 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5161 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5221 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5281 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5341 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5401 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5461 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5521 tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca
     5581 gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc
     5641 ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc
     5701 agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc
     5761 agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat
     5821 gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt
     5881 ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc
     5941 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
     6001 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
     6061 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
     6121 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
     6181 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
     6241 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
     6301 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
     6361 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
     6421 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
     6481 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
     6541 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     6601 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     6661 ccagtgcaag tgcaggtgcc agaacatttc tct
//
