LOCUS       Exported                7495 bp ds-DNA     circular SYN 12-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7495)
  AUTHORS   Noboru Mizushima
  TITLE     Direct Submission
  JOURNAL   Exported Oct 7, 2024 from SnapGene 4.0.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7495
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    381..676
                     /label=LTR
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1890..1913
                     /label=Universal Primer 1
     CDS             1995..2711
                     /codon_start=1
                     /label=Venus
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2030..2049)
                     /label=Universal Primer 13
     primer_bind     2654..2673
                     /label=Universal Primer 12
     primer_bind     2729..2757
                     /label=GS27_EcorI_Fw
     CDS             2739..3377
                     /codon_start=1
                     /label=GS27
                     /translation="MDPLFQQTHKQVHEIQSCMGRLETADKQSVHIVENEIQASIDQIF
                     SRLERLEILSSKEPPNKRQNAKLRVDQLKYDVQHLQTALRNFQHRRHAREQQERQREEL
                     LSRTFTTNDSDTTIPMDESLQFNSSLQKVHNGMDDLILDGHNILDGLRTQRLTLKGTQK
                     KILDIANMLGLSNTVMRLIEKRAFQDKYFMIGGMLLTCVVMFLVVQYLT"
     primer_bind     complement(3357..3377)
                     /label=GS27_XhoI_Rv
     misc_feature    3422..3974
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3438..3461)
                     /label=Universal Primer 2
     primer_bind     3900..3923
                     /label=Universal Primer 3
     CDS             3992..4591
                     /codon_start=1
                     /label=PuroR
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4233..4257)
                     /label=Universal Primer 7
     primer_bind     complement(4669..4694)
                     /label=Universal Primer 4
     LTR             4821..5290
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5349..5370)
                     /label=Universal Primer 6
     rep_origin      complement(5591..6179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6343..7203)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7420..7442
                     /label=Universal Primer 5
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 aaagcttggc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2041 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2101 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     2161 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     2221 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2281 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2341 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2401 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     2461 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     2521 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2581 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     2641 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2701 agctgtacaa gggttcaggc tcgggatccg ggaattcgat ggatcccctg ttccagcaaa
     2761 cgcacaagca ggtccacgag atccagtctt gcatgggacg cctggagacg gcagacaagc
     2821 agtctgtgca catagtagaa aacgaaatcc aagcaagcat agaccagata ttcagccgtc
     2881 tagaacgtct ggagattttg tccagcaagg agccccctaa caaaaggcaa aatgccaaac
     2941 ttcgggttga ccagttaaag tatgatgtcc agcacctgca gactgcgctc agaaacttcc
     3001 agcatcggcg ccatgcaagg gagcagcagg agagacagcg agaagagctt ctgtctcgaa
     3061 ccttcaccac taacgactct gacaccacca taccaatgga cgaatcactg cagtttaact
     3121 cctccctcca gaaagttcac aacggcatgg atgacctcat tttagatggg cacaatattt
     3181 tagatggact gaggacccag agactgacct tgaaggggac tcagaagaag atccttgaca
     3241 ttgccaacat gctgggcttg tccaacacag tgatgcggct catcgagaag cgggctttcc
     3301 aggacaagta ctttatgata ggtgggatgc tgctgacctg tgtggtcatg ttcctcgtgg
     3361 tgcagtacct gacatgaggg tctcgagagc ggccgctacg taaattccgc ccccccccct
     3421 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     3481 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3541 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3601 gtgaaggaag cagttcctct ggaagtttct tgaagacaaa caacgtctgt agcgaccctt
     3661 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3721 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3781 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3841 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3901 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3961 acacgatgat aatatggcca caaccgtcga catgaccgag tacaagccca cggtgcgcct
     4021 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     4081 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     4141 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4201 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4261 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4321 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4381 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4441 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4501 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4561 ctggtgcatg acccgcaagc ccggtgcctg agtcgaccgc ccgccccacg acccgcagcg
     4621 cccgaccgaa aggagcgcac gaccccatgg ctcgacgata aaataaaaga ttttatttag
     4681 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     4741 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     4801 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4861 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     4921 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     4981 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     5041 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     5101 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     5161 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     5221 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     5281 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca
     5341 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5401 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5461 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5521 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5581 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5641 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5761 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5941 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6001 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6061 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6121 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6181 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6241 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccggccg
     6301 caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6361 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6421 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6481 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6541 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6601 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6661 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6721 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6781 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6841 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6901 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     6961 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7201 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7261 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7321 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     7381 gcgtatcacg aggccctttc gtcttcacta taggagcttg gcccattgca tacgttgtat
     7441 ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc atgtt
//
