LOCUS       Exported                7514 bp ds-DNA     circular SYN 27-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Noboru Mizushima
  TITLE     Direct Submission
  JOURNAL   Exported Feb 27, 2024 from SnapGene 4.0.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7514
                     /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"
     primer_bind     1475..1498
                     /label=pLXSN 5'
     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=pBMN 5'
     primer_bind     1964..2017
                     /label=TSprimer57
     CDS             1973..3325
                     /codon_start=1
                     /label=MCS
                     /translation="IDIKLKGSMNLASQSGEAGAGQLLFANFNQDNTSLAVGSKSGYKF
                     FSLSSVDKLEQIYECTDTEDVCIVERLFSSSLVAIVSLKAPRKLKVCHFKKGTEICNYS
                     YSNTILAVKLNRQRLIVCLEESLYIHNIRDMKVLHTIRETPPNPAGLCALSINNDNCYL
                     AYPGSATIGEVQVFDTINLRAANMIPAHDSPLAALAFDASGTKLATASEKGTVIRVFSI
                     PEGQKLFEFRRGVKRCVSICSLAFSMDGMFLSASSNTETVHIFKLETVKEKPPEEPTTW
                     TGYFGKVLMASTSYLPSQVTEMFNQGRAFATVRLPFCGHKNICSLATIQKIPRLLVGAA
                     DGYLYMYNLDPQEGGECALMKQHRLDGSLETTNEILDSASHDCPLVTQTYGAAAGKGTY
                     VPSSPTRLAYTDDLGAVGGACLEDEASALRLDEDSEHPPMILRTDEFGSGSG"
     primer_bind     1991..2017
                     /label=TSprimer55
     misc_feature    1991..1996
                     /label=BamHI
     CDS             1997..3304
                     /codon_start=1
                     /label=MCS
                     /translation="MNLASQSGEAGAGQLLFANFNQDNTSLAVGSKSGYKFFSLSSVDK
                     LEQIYECTDTEDVCIVERLFSSSLVAIVSLKAPRKLKVCHFKKGTEICNYSYSNTILAV
                     KLNRQRLIVCLEESLYIHNIRDMKVLHTIRETPPNPAGLCALSINNDNCYLAYPGSATI
                     GEVQVFDTINLRAANMIPAHDSPLAALAFDASGTKLATASEKGTVIRVFSIPEGQKLFE
                     FRRGVKRCVSICSLAFSMDGMFLSASSNTETVHIFKLETVKEKPPEEPTTWTGYFGKVL
                     MASTSYLPSQVTEMFNQGRAFATVRLPFCGHKNICSLATIQKIPRLLVGAADGYLYMYN
                     LDPQEGGECALMKQHRLDGSLETTNEILDSASHDCPLVTQTYGAAAGKGTYVPSSPTRL
                     AYTDDLGAVGGACLEDEASALRLDEDSEHPPMILRTD"
     misc_feature    1997..3304
                     /label=WIPI2B CDS
     primer_bind     2056..2085
                     /label=TSprimer250
     primer_bind     complement(2056..2085)
                     /label=TSprimer251
     misc_feature    2071
                     /label=Tamura mutation 1
                     /note="g‚©?a‚É’uŠ·‚³‚ê‚Ä‚¢‚éD“¯‹`’uŠ·D"
     misc_feature    2215
                     /label=Tamura mutation 2
                     /note="c‚©?g‚É’uŠ·‚³‚ê‚Ä‚¢‚éD“¯‹`’uŠ·D"
     primer_bind     complement(3284..3337)
                     /label=TSprimer58
     primer_bind     complement(3287..3304)
                     /label=TSprimer56
     misc_feature    3305..3310
                     /label=EcoRI
     CDS             3326..3394
                     /codon_start=1
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3441..3993
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3774..3797)
                     /label=IRES Rev.
     primer_bind     3919..3942
                     /label=IRES Fwd.
     CDS             4011..4610
                     /codon_start=1
                     /label=PuroR
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4688..4713)
                     /label=Puro Rev.
     LTR             4840..5309
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(5610..6198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6362..7222)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7439..7461
                     /label=pBRforEco
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 aaagcttaaa ggatccatga acctggcgag ccagagcggg gaggccggcg ccggccagct
     2041 gctcttcgcc aacttcaacc aggacaacac gtccctagct gttggtagta agtccggtta
     2101 taaatttttc tccctttctt ctgtggataa gctggaacag atctatgaat gcaccgatac
     2161 ggaagatgtg tgcattgtag agagattgtt ctccagcagc ctagtggcca tcgtcagcct
     2221 taaagcacca aggaagctaa aggtttgcca ctttaagaag ggaactgaga tctgcaacta
     2281 cagctactcc aacacgattc tggctgtgaa gctcaacagg cagaggctga tagtatgcct
     2341 ggaggagtcc ctgtacatcc acaacattcg ggacatgaag gtgctgcata cgatcaggga
     2401 gacgcctcca aaccctgcag gcctgtgtgc gctgtcaatc aacaacgaca actgctactt
     2461 ggcgtaccca gggagcgcga ccatcggaga ggtgcaggtc ttcgatacca ttaatttgag
     2521 agctgcaaac atgattccgg ctcacgacag tcctttagcg gcactggcct ttgacgcaag
     2581 tggaactaaa cttgccacgg cttcggagaa ggggaccgtg attagggtat tttccattcc
     2641 agaaggacaa aaactctttg agtttcggag aggagtaaag aggtgcgtga gcatctgctc
     2701 cctggccttc agcatggacg gcatgttcct ctccgcctcc agcaacactg agaccgtgca
     2761 catcttcaaa ctcgagactg tgaaagaaaa acccccagag gagcccacca cctggaccgg
     2821 gtacttcggg aaagtgctca tggcctccac cagctacctg ccttcccaag tgacagaaat
     2881 gttcaaccag ggcagagcct tcgccacggt ccgcctgcca ttctgcggcc acaaaaacat
     2941 ctgctcgcta gccacaattc agaagatccc gcggttgttg gtgggtgccg ccgacgggta
     3001 cctgtacatg tacaacctgg acccccagga gggcggcgag tgtgccctga tgaagcagca
     3061 ccggctggac ggcagtctgg aaacgaccaa tgagatcttg gactctgcct ctcacgactg
     3121 ccccttagtc actcagacat acggcgcagc tgcaggaaaa ggtacttacg tgccttcatc
     3181 cccaacgaga cttgcctaca cagacgacct gggtgctgtg ggtggcgcct gcctggagga
     3241 cgaggccagc gccctgcgcc tggatgagga cagcgagcac ccgcccatga ttcttcggac
     3301 tgacgaattc ggctctggtt caggcgacta caaagaccat gacggtgatt ataaagatca
     3361 tgacatcgac tacaaggatg acgatgacaa gtaatgattt ctcgagagcg gccgctacgt
     3421 aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     3481 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     3541 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     3601 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagtttctt gaagacaaac
     3661 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     3721 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     3781 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     3841 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     3901 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     3961 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccgtcgac atgaccgagt
     4021 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg
     4081 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg
     4141 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4201 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4261 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4321 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4381 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4441 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4501 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4561 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgaccgcc
     4621 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc tcgacgataa
     4681 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4741 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     4801 tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
     4861 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     4921 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     4981 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     5041 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     5101 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     5161 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     5221 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     5281 attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt
     5341 tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg
     5401 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5461 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5521 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5581 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5641 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5701 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5761 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5821 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5881 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5941 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6001 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6061 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6121 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6181 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6241 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6301 atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6961 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7381 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcactat aggagcttgg
     7441 cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     7501 attaccgcca tgtt
//
