LOCUS       40924_9771        7528 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vivid Colors(TM) Gateway(R) destination vector for expressing 
            N-terminally Emerald GFP-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7528)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7528)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             813..1529
                     /codon_start=1
                     /label=EmGFP
                     /note="Emerald GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    1545..1669
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1694..1724
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1778..2434
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2779..3081
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3125..3249)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3257..3298
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3382..3430
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3632..4058
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4072..4401
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        4449..4496
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4515..4910
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5071..5204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5241..5257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5265..5281)
                     /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(5289..5319)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5334..5355)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5643..6231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6405..7262)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7263..7367)
                     /label=AmpR promoter
ORIGIN
        1 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
       61 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      721 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      781 agaggatccc taccggtgat atcctcgaga ccatggtgag caagggcgag gagctgttca
      841 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      901 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      961 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcacc tacggcgtgc
     1021 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1081 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1141 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1201 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1261 aggtctatat caccgccgac aagcagaaga acggcatcaa ggtgaacttc aagacccgcc
     1321 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1381 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1441 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1501 tcactctcgg catggacgag ctgtacaagg gctcgagccc atcaacaagt ttgtacaaaa
     1561 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     1621 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt
     1681 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
     1741 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     1801 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     1861 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     1921 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     1981 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     2041 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     2101 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     2161 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     2221 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     2281 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     2341 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     2401 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaaga tctggatccg gcttactaaa
     2461 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     2521 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     2581 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     2641 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     2701 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     2761 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     2821 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     2881 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     2941 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     3001 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     3061 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     3121 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     3181 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     3241 caaagtggtt gataacggga agcctatccc taaccctctc ctcggtctcg attctacgcg
     3301 taccggttag taatgagttt aaacggggga ggctaactga aacacggaag gagacaatac
     3361 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     3421 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3481 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3541 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     3601 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttcccgt caagctctaa
     3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaact
     3841 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3901 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3961 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4021 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4081 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4261 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4321 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4381 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4441 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4501 tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     4561 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     4621 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     4681 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     4741 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     4801 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     4861 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacttc
     4921 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     4981 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5041 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5101 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5161 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5221 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5281 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5341 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5401 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5461 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5521 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5581 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5641 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5701 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5761 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5821 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5881 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5941 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6001 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6061 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6241 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6301 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6361 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6421 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6481 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6541 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6601 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6661 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6721 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6781 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6841 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6901 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6961 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7021 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7081 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7141 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7201 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7261 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7321 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7381 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     7441 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7501 aggtcgctga gtagtgcgcg agcaaaat
//
