LOCUS       40924_9746        7442 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vivid Colors(TM) Gateway(R) destination vector for expressing 
            C-terminally Emerald GFP-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7442)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7442)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7442
                     /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"
     promoter        733..751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    785..909
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        934..964
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1018..1674
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2019..2321
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2365..2489)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2503..3219
                     /codon_start=1
                     /label=EmGFP
                     /note="Emerald GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3294..3342
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3544..3972
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3986..4315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        4363..4410
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4429..4824
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4985..5118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5155..5171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5179..5195)
                     /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(5203..5233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5248..5269)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5557..6145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6319..7176)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7177..7281)
                     /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 gcagagctct ctggctaact agagaaccca ctgcttactg
      721 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt taagctgagc
      781 atcaacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
      841 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
      901 gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      961 aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg
     1021 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1081 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1141 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1201 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     1261 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1321 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1381 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1441 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     1501 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1561 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1621 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaaga
     1681 tctggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
     1741 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga
     1801 agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga
     1861 tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc
     1921 cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat
     1981 gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct
     2041 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
     2101 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc
     2161 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
     2221 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
     2281 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta
     2341 tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat
     2401 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     2461 tctcgttcag ctttcttgta caaagtggtt gatgctgtta acatggtgag caagggcgag
     2521 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2581 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2641 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcacc
     2701 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2761 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2821 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2881 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2941 aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa ggtgaacttc
     3001 aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3061 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3121 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3181 gccgccggga tcactctcgg catggacgag ctgtacaagt aatgataagt ttaaacgggg
     3241 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3301 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3361 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3421 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3481 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     3541 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3601 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3661 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3721 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3781 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3841 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3901 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3961 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4021 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4081 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4141 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4201 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4261 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4321 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     4381 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct
     4441 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     4501 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     4561 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     4621 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     4681 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     4741 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     4801 ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg actgacacgt
     4861 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4921 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4981 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5101 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5161 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5221 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5281 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6301 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6361 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6421 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6481 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6541 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6601 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6661 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6721 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6781 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6841 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6901 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6961 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7021 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7081 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7141 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7321 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7381 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7441 at
//
