LOCUS       Exported                6909 bp DNA     circular SYN 02-JUL-2018
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6909)
  AUTHORS   Noboru Mizushima
  TITLE     Direct Submission
  JOURNAL   Exported May 27, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    381..676
                     /label=LTR
                     /note="color: #ffe4c4"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
                     /note="color: #ffe4c4"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
                     /note="color: #ffff99; direction: RIGHT"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
                     /note="color: #ffe4c4
                     Cleavage site after base 1941"
     primer_bind     1890..1913
                     /label=Universal Primer 1
                     /note="color: black; sequence: GGCATCGCAGCTTGGATACACGCC"
     CDS             1994..2710
                     /codon_start=1
                     /label=EGFP
                     /note="color: #ccffcc"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2029..2048)
                     /label=Universal Primer 13
                     /note="color: black; sequence: GCTCGACCAGGATGGGCACC"
     primer_bind     2653..2672
                     /label=Universal Primer 12
                     /note="color: black; sequence: CCTGCTGGAGTTCGTGACCG"
     primer_bind     complement(2717..2811)
                     /label=278_Gb_IPUG-Ec-8KFarn-Xh_r
                     /note="color: black; sequence: 
                     aGCGGCCGCtCTCGAGttaCATAATCACGCATTTGGTTTTACTTTTCTTCTTCTTTTTT
                     TTTCCATCTTTGGATGAATTCccGGATCCcgagcc"
     primer_bind     2717..2811
                     /label=277_Gb_IPUG-Ec-8KFarn-Xh_f
                     /note="color: black; sequence: 
                     ggctcgGGATCCggGAATTCATCCAAAGATGGAAAAAAAAAGAAGAAGAAAAGTAAAAC
                     CAAATGCGTGATTATGtaaCTCGAGaGCGGCCGCt"
     misc_feature    2731..2736
                     /label=EcoRI
                     /note="color: #cc99ff"
     misc_feature    2738..2791
                     /label=8K-Farn
                     /note="color: #ffcc00"
     misc_feature    2796..2801
                     /label=XhoI
                     /note="color: #cc99ff"
     primer_bind     complement(2802..2811)
                     /label=277_Gb_IPUG-Ec-8KFarn-Xh_f
                     /note="color: black; sequence: 
                     ggctcgGGATCCggGAATTCATCCAAAGATGGAAAAAAAAAGAAGAAGAAAAGTAAAAC
                     CAAATGCGTGATTATGtaaCTCGAGaGCGGCCGCt"
     misc_feature    2836..3388
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(2852..2875)
                     /label=Universal Primer 2
                     /note="color: black; sequence: GCACACCGGCCTTATTCCAAGCGG"
     primer_bind     3314..3337
                     /label=Universal Primer 3
                     /note="color: black; sequence: GTCGAGGTTAAAAAAACGTCTAGG"
     misc_feature    3406..4005
                     /label=PuroR
                     /note="color: #ccffcc; direction: RIGHT"
     primer_bind     complement(3647..3671)
                     /label=Universal Primer 7
                     /note="color: black; sequence: GCGAACACCGCCCCCGCTTCGACGC"
     primer_bind     complement(4083..4108)
                     /label=Universal Primer 4
                     /note="color: black; sequence: CCTTTTTCTGGAGACTAAATAAAATC"
     LTR             4235..4704
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
                     /note="color: #ffe4c4"
     primer_bind     complement(4763..4784)
                     /label=Universal Primer 6
                     /note="color: black; sequence: CGATTCATTAATGCAACTATGG"
     rep_origin      complement(5005..5593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff99"
     CDS             complement(5757..6617)
                     /codon_start=1
                     /label=AmpR
                     /note="color: #ccffcc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6834..6856
                     /label=Universal Primer 5
                     /note="color: black; sequence: GGCCCATTGCATACGTTGTATCC"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat
     1981 aaagcttgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2041 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2101 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2161 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2221 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2281 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2341 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2401 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2461 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2521 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2581 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2641 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2701 gctgtacaag ggttcaggct cgggatccgg gaattcatcc aaagatggaa aaaaaaagaa
     2761 gaagaaaagt aaaaccaaat gcgtgattat gtaaactcga gagcggccgc tacgtaaatt
     2821 ccgccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2881 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2941 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3001 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagt ttcttgaaga caaacaacgt
     3061 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     3121 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     3181 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3241 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     3301 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3361 ttttcctttg aaaaacacga tgataatatg gccacaaccg tcgacatgac cgagtacaag
     3421 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3481 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3541 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3601 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3661 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3721 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3781 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3841 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3901 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     3961 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga ccgcccgccc
     4021 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctcgac gataaaataa
     4081 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     4141 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     4201 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     4261 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     4321 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4381 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4441 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4501 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4561 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     4621 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     4681 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     4741 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     4801 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4861 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4921 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4981 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5041 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5101 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5161 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5221 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5281 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5341 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5401 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5461 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5521 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5581 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5641 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5701 tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5761 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5821 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5881 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5941 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6001 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6061 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6121 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6181 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6241 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6301 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6361 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6421 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6481 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6541 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6601 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6661 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6721 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6781 acctataaaa ataggcgtat cacgaggccc tttcgtcttc actataggag cttggcccat
     6841 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
     6901 cgccatgtt
//
