LOCUS       pCMV-Hs.cargo(Cas9)               10649 bp DNA     circular SYN 05-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10649)
  AUTHORS   Segel, M., Lash, B., Song, J., Ladha, A., Liu, C.C., Jin, X., Mekhedov, S., Macrae, R.K., Koonin, E.V., Zhang, F. 
  TITLE     Mammalian retrovirus-like protein PEG10 packages its own mRNA and can be pseudotyped for mRNA delivery
  JOURNAL   Science
FEATURES             Location/Qualifiers
     source          1..10649
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        236..852
     regulatory      315..602
                     /standard_name="CAG_enhancer"
                     /label=CAG_enhancer
     promoter        863..881
     gene            929..1407
                     /standard_name="PEG10"
                     /label=PEG10
     exon            929..1184
                     /standard_name="PEG10-001 Exon 1"
                     /label=PEG10-001 Exon 1
     exon            1185..1407
                     /standard_name="PEG10-001 Exon 2"
                     /label=PEG10-001 Exon 2
     CDS             1417..1443
                     /standard_name="HA"
                     /label=HA
     CDS             1447..1467
                     /standard_name="SV40 NLS"
                     /label=SV40 NLS
     CDS             1477..5577
                     /standard_name="Cas9"
                     /label=Cas9
     CDS             5581..5601
                     /standard_name="SV40 NLS"
                     /label=SV40 NLS
     exon            5614..6128
                     /standard_name="PEG10-001 Exon 2"
                     /label=PEG10-001 Exon 2
     gene            5614..6128
                     /standard_name="PEG10"
                     /label=PEG10
     misc_feature    complement(6243..6260)
                     /standard_name="BGH_rev_primer"
                     /label=BGH_rev_primer
                     /note="Geneious type: primer"
     terminator      6246..6473
     misc_feature    complement(6956..6976)
                     /standard_name="pBABE_3_primer"
                     /label=pBABE_3_primer
                     /note="Geneious type: primer"
     regulatory      complement(6962..7177)
                     /standard_name="SV40_enhancer"
                     /label=SV40_enhancer
     promoter        6974..7242
     misc_feature    7203..7222
                     /standard_name="SV40pro_F_primer"
                     /label=SV40pro_F_primer
                     /note="Geneious type: primer"
     gene            7360..8148
                     /standard_name="NeoR/KanR"
                     /label=NeoR/KanR
     terminator      8328..8447
     misc_feature    8416..8435
                     /standard_name="EBV_rev_primer"
                     /label=EBV_rev_primer
                     /note="Geneious type: primer"
     promoter        complement(8491..8509)
     misc_feature    complement(8508..8530)
                     /standard_name="M13_pUC_rev_primer"
                     /label=M13_pUC_rev_primer
                     /note="Geneious type: primer"
     promoter        complement(8544..8573)
     gene            complement(9653..10513)
                     /standard_name="Ampicillin"
                     /label=Ampicillin
     promoter        complement(10555..10583)
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagataata cgactcacta
      901 tagcccaagc tggctagcgt ttaaaaccct cctcggtgca acctatataa ggctcacagt
      961 ctgcgctcct ggtacacgcg cttcaacttc ggttggtgtg tgtcgaagaa acctgactgc
     1021 gccctgagga gaacagcgga gaaggtccac cgagcctggc gaaaggtccg ctgagcgggc
     1081 tgtcgtccgg agccactccg ggctgcggag cacccagtgg agaccgcgcc tggctcaggt
     1141 gtgggacccc atccttcctg tcttcgcaga ggagtcctcg cgtgaaataa gcgggttttg
     1201 aaaacaaaaa aaagaaggag tggaagaggg ggccaggatc caggcctcca tccccacaga
     1261 agtgaagcta cagctgggag gtctcctccc accccaaccg tcaccctggg tcccgactgc
     1321 ccacctcctc ctcctccccc tccccccaac aacaacaaca acaacaactc caagcacacc
     1381 ggccataaga gtgcgtgtgt ccccaacgcc accatgtacc catacgatgt tccagattac
     1441 gcttcgccga agaaaaagcg caaggtcgaa gcgtccgaca agaagtacag catcggcctg
     1501 gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     1561 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     1621 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     1681 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     1741 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     1801 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     1861 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     1921 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     1981 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     2041 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     2101 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     2161 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     2221 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     2281 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     2341 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     2401 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     2461 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     2521 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     2581 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     2641 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     2701 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     2761 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     2821 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     2881 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     2941 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     3001 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     3061 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     3121 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     3181 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     3241 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     3301 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     3361 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     3421 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     3481 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     3541 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     3601 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     3661 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     3721 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     3781 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     3841 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     3901 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     3961 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     4021 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     4081 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     4141 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     4201 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     4261 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     4321 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     4381 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     4441 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     4501 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     4561 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     4621 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     4681 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     4741 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     4801 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     4861 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     4921 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     4981 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     5041 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     5101 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     5161 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     5221 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     5281 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     5341 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     5401 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     5461 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     5521 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacagc
     5581 cccaagaaga agagaaaggt ggaggccagc taaatacctg tcatgtcctt caggatctct
     5641 gccctcaaaa tttattcctg ttcagcttct caatcagtga ctgtgtgcta aattttaggc
     5701 tactgtatct tcaggccacc tgaggcacat cctctctgaa acggctatgg aaggttaggg
     5761 ccactctgga ctggcacaca tcctaaagca ccaaaagacc ttcaacattt tctgagagca
     5821 acagagtatt tgccaataaa tgatctctca tttttccacc ttgactgcca atctaactaa
     5881 aataattaat aagtttactt tccagccagt cctggaagtc tgggttttac ctgccaaaac
     5941 ctccatcacc atctaaatta taggctgcca aatttgctgt ttaacattta cagagaagct
     6001 gatacaaacg caggaaatgc tgatttcttt atggaggggg agacgaggag gaggaggaca
     6061 tgacttttct tgcggtttcg gtaccctctt tttaaatcac tggaggactg aggccttatt
     6121 aaggaagcct taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg
     6181 cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc
     6241 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6301 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6361 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     6421 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     6481 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     6541 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     6601 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6661 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6721 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6781 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6841 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6901 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     6961 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     7021 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     7081 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     7141 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     7201 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     7261 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     7321 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     7381 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     7441 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     7501 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     7561 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     7621 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     7681 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     7741 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7801 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7861 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7921 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7981 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     8041 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     8101 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     8161 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     8221 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     8281 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     8341 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     8401 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     8461 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     8521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     8581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     8641 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     8701 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     8761 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     8821 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     8881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     8941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9061 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     9241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     9301 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     9361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     9421 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9481 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9541 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     9601 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9661 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9721 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9781 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9841 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9901 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9961 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    10021 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    10081 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    10141 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    10201 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    10261 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    10321 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    10381 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10441 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    10501 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    10561 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    10621 catttccccg aaaagtgcca cctgacgtc
//
