LOCUS       Exported                6978 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pEGFPC4 converted Gateway Destination vector for expressing EGFP 
            tagged fusion proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pEZYegfp
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  AUTHORS   Guo F, Chiang MY, Wang Y, Zhang YZ
  TITLE     An in vitro recombination method to convert restriction- and 
            ligation-independent expression vectors.
  JOURNAL   Biotechnol J. 2008 Mar . 3(3):370-7.
  PUBMED    18064608
REFERENCE   2  (bases 1 to 6978)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6978
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             613..1329
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1360..1484
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1913..2218)
                     /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     complement(2015..2034)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2568..3221)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     primer_bind     3136..3155
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3222..3324)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3502..3626)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3766..3887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3803..3822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3857..3876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3894..4349)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4031..4052)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4243..4262
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4376..4480
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4482..4839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4505..4525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4690..4825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4752..4771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4874..5668
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4928..4947)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5538..5557
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5856..5875)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5900..5947
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6276..6864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6765..6784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctgcagaa ttgattaata caagtttgta caaaaaagct
     1381 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
     1441 cagactacat aatactgtaa aacacaacat atccagtcac tatgaatcaa ctacttagat
     1501 ggtattagtg acctgtagtc gaccgacagc cttccaaatg ttcttcgggt gatgctgcca
     1561 acttagtcga ccgacagcct tccaaatgtt cttctcaaac ggaatcgtcg tatccagcct
     1621 actcgctatt gtcctcaatg ccgtattaaa tcataaaaag aaataagaaa aagaggtgcg
     1681 agcctctttt ttgtgtgaca aaataaaaac atctacctat tcatatacgc tagtgtcata
     1741 gtcctgaaaa tcatctgcat caagaacaat ttcacaactc ttatactttt ctcttacaag
     1801 tcgttcggct tcatctggat tttcagcctc tatacttact aaacgtgata aagtttctgt
     1861 aatttctact gtatcgacct gcagactggc tgtgtataag ggagcctgac atttatattc
     1921 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
     1981 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc
     2041 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
     2101 gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta
     2161 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt
     2221 caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca
     2281 gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc
     2341 tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat
     2401 agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat
     2461 acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc
     2521 atactgttat ctggctttta gtaagccgga tccacgcggc gtttacgccc cccctgccac
     2581 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     2641 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     2701 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     2761 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     2821 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     2881 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     2941 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     3001 cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     3061 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     3121 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     3181 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     3241 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     3301 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc
     3361 ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt
     3421 atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tagtcgacta caggtcacta
     3481 ataccatcta agtagttgat tcatagtgac tggatatgtt gtgttttaca gtattatgta
     3541 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     3601 cgttcagctt tcttgtacaa agtggtcctc gagctcggat ccaccggatc tagataactg
     3661 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3721 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3901 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3961 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4021 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4081 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4141 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4201 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4261 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4321 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4381 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4441 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4501 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4561 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4681 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4741 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4801 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4861 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4921 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4981 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5041 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5101 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5161 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5221 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5281 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5341 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5401 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5461 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5521 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5581 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5641 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5701 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5761 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5821 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5881 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5941 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     6001 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     6061 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6121 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6181 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6241 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6301 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6361 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6421 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6481 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6541 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6601 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6661 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6721 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6781 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6841 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6901 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6961 cgtattaccg ccatgcat
//
