LOCUS       sequence-202612-        6830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6830)
  AUTHORS   Lu G, Middleton RE, Sun H, Naniong M, Ott CJ, Mitsiades CS, Wong KK,
            Bradner JE, Kaelin WG Jr
  TITLE     The myeloma drug lenalidomide promotes the cereblon-dependent 
            destruction of Ikaros proteins.
  JOURNAL   Science. 2014 Jan 17;343(6168):305-9. doi: 10.1126/science.1244917. 
            Epub 2013 Nov 29.
  PUBMED    24292623
REFERENCE   2  (bases 1 to 6830)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.1244917"; journalName: "Science"; date: 
            "2014-01-17- 17"; volume: "343"; issue: "6168"; pages: "305-9"
COMMENT     Sequence Label: pcDNA3-FLAG- gate-pGK-HYG
FEATURES             Location/Qualifiers
     source          1..6830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..138
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(163..168,169..186,187..193)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             247..906
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(313..332)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1248..1553
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1432..1451
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1594..1718)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1862..2361
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1897..1920)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2294..2313
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2379..3404
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2379..3404
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    3534..3655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3571..3590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3625..3644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3704..3720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3704..3720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3717..3739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3728..3744
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(3752..3758,3759..3776,3777..3782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3797..3818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3935..3952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4106..4694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4186..4205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4865..5656,5657..5725))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5488..5507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5726..5830)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5905..5924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6096..6475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6476..6679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6630..6650
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6724..6743
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6724..6742
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6778..6787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6787..6810
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tcgaggttat caaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
       61 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
      121 acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc
      181 ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa
      241 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
      301 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
      361 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
      421 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
      481 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
      541 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
      601 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
      661 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
      721 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
      781 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
      841 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
      901 gcgtaaacgc gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
      961 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
     1021 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     1081 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
     1141 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     1201 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
     1261 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     1321 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     1381 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     1441 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
     1501 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
     1561 gctcccttat acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta
     1621 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     1681 ttttacgttt ctcgttcagc tttcttgtac aaagtggttc gataacgaat tccgcccccc
     1741 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     1801 ttattttcca ccatattgcc gtcttttggc aatgtgaggg ccgcgtttaa cgaattctac
     1861 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
     1921 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
     1981 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
     2041 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
     2101 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
     2161 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
     2221 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
     2281 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
     2341 tcctcatctc cgggcctttc gacctgcatc ccgccaccat gaaaaagcct gaactcaccg
     2401 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     2461 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     2521 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     2581 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     2641 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     2701 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     2761 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     2821 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     2881 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     2941 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     3001 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     3061 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     3121 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     3181 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     3241 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     3301 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     3361 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
     3421 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3481 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3541 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3601 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3661 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3721 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3781 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3841 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3901 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5881 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5941 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     6001 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     6061 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     6121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     6481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     6661 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     6721 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagcttgcc
     6781 accatggact acaaggacga cgatgacaag ctcgatggag gtgcggccgc
//
