LOCUS       40924_25666        5307 bp DNA     circular SYN 01-JAN-1980
DEFINITION  IRES-containing bicistronic vector for expressing a gene together 
            with the fluorescent protein AcGFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5307)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5307)
  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..5307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     misc_feature    600..665
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    667..1253
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1254..1970
                     /codon_start=1
                     /label=AcGFP1
                     /note="Aequorea coerulescens GFP"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2095..2216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2223..2678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2705..2809
                     /label=AmpR promoter
     promoter        2811..3168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3203..3994
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4229..4276
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4605..5193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
      721 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
      781 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
      841 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
      901 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
      961 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     1021 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     1081 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     1141 tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     1201 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga
     1261 gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg aatggcgatg
     1321 tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc tacggcaagc
     1381 tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc accctggtga
     1441 ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg aagcagcacg
     1501 acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc ttcttcgagg
     1561 atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc ctggtgaatc
     1621 gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc aataagatgg
     1681 agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag aatggcatca
     1741 aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg gccgaccact
     1801 accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac cactacctgt
     1861 ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg atctacttcg
     1921 gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag tgagcggccg
     1981 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2041 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2101 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2161 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2221 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2281 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2341 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2401 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2461 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2521 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2581 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2641 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2701 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2761 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2821 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3001 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3061 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3121 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3181 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3241 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3301 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3361 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3421 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3481 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3541 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3601 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3661 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3721 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3781 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3841 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3901 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3961 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4021 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4081 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4141 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4201 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4261 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4321 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4381 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4441 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4501 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4561 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4621 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4681 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4741 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4801 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4861 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4921 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4981 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5041 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5101 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5161 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5221 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5281 gtggataacc gtattaccgc catgcat
//
