LOCUS       40924_16905        6040 bp DNA     circular SYN 01-JAN-1980
DEFINITION  IRES-containing bicistronic vector for expressing a gene together 
            with the ZsGreen1 fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6040)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6040)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The gene inserted into the MCS should contain start and stop codons.
FEATURES             Location/Qualifiers
     source          1..6040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        165..1346
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_feature    1357..1422
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1424..2010
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2011..2703
                     /codon_start=1
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    2828..2949
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2956..3411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3438..3542
                     /label=AmpR promoter
     promoter        3544..3901
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3936..4727
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4962..5009
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5338..5926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tgagtaattc atacaaaagg actcgcccct gccttgggga atcccaggga
       61 ccgtcgttaa actcccacta acgtagaacc cagagatcgc tgcgttcccg ccccctcacc
      121 cgcccgctct cgtcatcact gaggtggaga agagcatgcg tgaggctccg gtgcccgtca
      181 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      241 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      301 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      361 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      421 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cgcccctggc
      481 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      541 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggctt gggcgctggg
      601 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      661 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      721 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      781 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
      841 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
      901 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
      961 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     1021 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1081 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1141 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1201 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1261 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1321 tttttcttcc atttcaggtg tcgtgacgct agcgctaccg gactcagatc tcgagctcaa
     1381 gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccgcccctct ccctcccccc
     1441 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     1501 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     1561 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     1621 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     1681 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     1741 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     1801 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     1861 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc tttacatgtg
     1921 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     1981 gaaaaacacg atgataatat ggccacaacc atggcccagt ccaagcacgg cctgaccaag
     2041 gagatgacca tgaagtaccg catggagggc tgcgtggacg gccacaagtt cgtgatcacc
     2101 ggcgagggca tcggctaccc cttcaagggc aagcaggcca tcaacctgtg cgtggtggag
     2161 ggcggcccct tgcccttcgc cgaggacatc ttgtccgccg ccttcatgta cggcaaccgc
     2221 gtgttcaccg agtaccccca ggacatcgtc gactacttca agaactcctg ccccgccggc
     2281 tacacctggg accgctcctt cctgttcgag gacggcgccg tgtgcatctg caacgccgac
     2341 atcaccgtga gcgtggagga gaactgcatg taccacgagt ccaagttcta cggcgtgaac
     2401 ttccccgccg acggccccgt gatgaagaag atgaccgaca actgggagcc ctcctgcgag
     2461 aagatcatcc ccgtgcccaa gcagggcatc ttgaagggcg acgtgagcat gtacctgctg
     2521 ctgaaggacg gtggccgctt gcgctgccag ttcgacaccg tgtacaaggc caagtccgtg
     2581 ccccgcaaga tgcccgactg gcacttcatc cagcacaagc tgacccgcga ggaccgcagc
     2641 gacgccaaga accagaagtg gcacctgacc gagcacgcca tcgcctccgg ctccgccttg
     2701 ccctgagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2761 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2821 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2881 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2941 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3001 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3061 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3121 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3181 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3241 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3301 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3361 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3421 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3481 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3541 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3841 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3901 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3961 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4021 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4081 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4141 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4201 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4261 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4321 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4381 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4441 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4501 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4561 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4621 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4681 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4741 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4801 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4861 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4921 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4981 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5041 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5101 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5161 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5221 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5281 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5821 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5881 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5941 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6001 atcccctgat tctgtggata accgtattac cgccatgcat
//
