LOCUS       62056_2225        7057 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..515
                     /label=MSCV
                     /note="Murine embryonic stem cell virus promoter including
                     the enhancer and promoter region of PCMV virus and 5' 
                     untranslated region of dl-587rev retrovirus"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             987..1403
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     CDS             1425..1448
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             1461..1487
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    complement(1981..2005)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2022..2597
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2598..3314
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     LTR             3419..3933
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     primer_bind     complement(4102..4118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4126..4142)
                     /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(4150..4180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4195..4216)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4504..5092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5266..6123)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6124..6228)
                     /label=AmpR promoter
     LTR             7056..7057
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctgccac catggactac aaggatgacg
     1441 atgacaagct cgatggagga tacccatacg atgttccaga ttacgctgct cgaggttatc
     1501 aaacaagttt gtacaaaaaa gttggctttc aggacccaca ggagcgaccc agaaagttac
     1561 cacagttatg cacagagctg caaacaacta tacatgatat aatattagaa tgtgtgtact
     1621 gcaagcaaca gttactgcga cgtgaggtat atgactttgc ttttcgggat ttatgtatag
     1681 tatatagaga tgggaatcca tatgctgtat gtgataaatg tttaaagttt tattctaaaa
     1741 ttagtgagta tagacattat tgttatagtg tgtatggaac aacattagaa cagcaataca
     1801 acaaaccgtt gtgtgatttg ttaattaggt gtattaactg tcaaaagcca ctgtgtcctg
     1861 aagaaaagca aagacatctg gacaaaaagc aaagattcca taatataagg ggtcggtgga
     1921 ccggtcgatg tatgtcttgt tgcagatcat caagaacacg tagagaaacc cagctgtaat
     1981 acccaacttt cttgtacaaa gtggttcgat aacgaattcc gccccccccc cctaacgtta
     2041 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     2101 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     2161 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2221 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2281 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2341 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2401 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2461 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     2521 taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     2581 ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2641 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2701 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2761 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2821 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2881 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2941 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3001 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3061 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3121 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3181 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3241 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3301 acgagctgta caagtaatga attaattaag aattatcaag cttatcgata ccgtcgacct
     3361 gcagccaagc ttatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3421 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3481 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     3541 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3601 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3661 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3721 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3781 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     3841 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3901 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     3961 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     4021 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aagagctggg
     4081 gagaattaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4141 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4201 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4261 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4321 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4381 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4441 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4501 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4561 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4621 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4681 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4741 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4801 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4861 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4921 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4981 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5041 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5101 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5161 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5221 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5281 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5341 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5401 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5461 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5521 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5581 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5641 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5701 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5761 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5821 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5881 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5941 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6001 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6061 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6121 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6181 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6241 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6301 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6361 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6421 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6481 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6541 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6601 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6661 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg
     6721 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     6781 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     6841 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     6901 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
     6961 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     7021 taaaagattt tatttagtct ccagaaaaag gggggaa
//
