LOCUS       pPU-Empty-miR30-L1221        6570 bp ds-DNA     circular SYN 29-AUG-2019
DEFINITION  Final.
ACCESSION   urn.local...hv-aw3pfuq
VERSION     .
KEYWORDS    pPU-HA.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7888)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tue, Feb 10, 2015 from SnapGene Viewer 2.6.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     rep_origin      236..579
                     /note="SV40ori"
                     /note="SV40 ori/prom"
     promoter        580..1161
                     /note="CMVp"
                     /note="CMVieProm"
                     /modified_by="User"
                     /label="CMV promoter"
     5'UTR           1180..1276
                     /note="5'R"
                     /note="5' U3"
     5'UTR           1277..1361
                     /note="5'U5"
                     /note="5' U5"
     CDS             1515..1882
                     /codon_start=1
                     /note="gag"
                     /translation="MGARASVLSGGELDRDGKKFG*GQGERKNIN*NI*YGQAGS*ND
                     SQLILAC*KHQKAVDKYWDSYNHPFRQDQKNLDHYIIQ*QPSIVCIKG*R*KTPRKL*
                     TR*RKSKTKVRPPHSKRPAA"
     CDS             1883..2738
                     /codon_start=1
                     /note="env"
                     /note="env sequence"
                     /translation="SSDLEEEI*GTIGEVNYINIK**KLNH*E*HPPRQREEWCREKK
                     EQWE*ELCSLGSWEQQEALWAQRQ*R*RYRPDNYCLV*CSSRTIC*GLLRRNSICCNS
                     QSGASSSSRQESWLWKDT*RINSSWGFGVALENSFAPLLCLGMLVGVINLWNRFGITR
                     PGWSGTEKLTITQA*YTP*LKNRKTSKKRMNKNYWN*INGQVCGIGLT*QIGCGI*NY
                     S****EAW*V*E*FLLYFL**IELGRDIHHYRFRPTSQPRGDPTGPKE*KKKVERETE
                     TDPFD**TD"
     misc_signal     2026..2265
                     /note="RRE"
     misc_feature    2764..2941
                     /created_by="User"
                     /label="Charneau"
     misc_signal     2792..2807
                     /note="cPPT"
     misc_signal     2871..2911
                     /note="CTS"
                     /note="Charneau Central Termination Signal"
     promoter        2950..3451
                     /note="SFFVp"
                     /note="SFFVprom"
                     /modified_by="User"
                     /label="SFFV promoter"
     CDS             3464..4060
                     /codon_start=1
                     /note="puro"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
                     LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
                     ETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
                     /modified_by="User"
                     /label="PuroR"
     CDS             4061..4288
                     /codon_start=1
                     /note="ubi K48R"
                     /note="Ubi K48R"
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLI
                     FAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
                     /modified_by="User"
                     /label="Ub K48R"
     modified_base   4202..4204
                     /created_by="User"
                     /label="K48R"
     CDS             4289..4321
                     /codon_start=1
                     /note="HA tag"
                     /translation="MGYPYDVPDYA"
                     /modified_by="User"
                     /label="HA tag"
     misc_feature    4342..4494
                     /ApEinfo_fwdcolor="#0080c0"
                     /ApEinfo_revcolor="#80ffff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="miR30"
     misc_signal     4520..4535
                     /note="PPT"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="PPT"
     3'UTR           4535..4559
                     /created_by="User"
                     /label="3' deltaU3"
     3'UTR           4568..4664
                     /note="3'R"
                     /note="3' R"
     3'UTR           4665..4748
                     /note="3'U5"
                     /note="3' U5"
     oriT            4759..5429
                     /note="colE1"
     marker          complement(5577..6437)
                     /created_by="User"
                     /label="Ampicillin marker"
     promoter        complement(6479..6507)
                     /created_by="User"
                     /label="Amp promoter"
ORIGIN      
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgctgtg
      241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      541 aggaggcttt tttggaggcc taggcttttg caaaaagctt tgacattgat tattgactag
      601 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      661 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      721 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      781 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      841 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      901 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      961 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1021 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1081 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1141 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
     1201 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1261 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     1321 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     1381 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     1441 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     1501 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     1561 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     1621 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1681 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1741 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1801 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1861 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1921 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1981 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     2041 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     2101 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     2161 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     2221 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     2281 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     2341 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     2401 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     2461 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     2521 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     2581 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     2641 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     2701 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
     2761 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     2821 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     2881 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     2941 gttaattaac tgcagccccg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3001 gaatgaaaga ccccacctgt aggtttggca agctagctgc agtaacgcca ttttgcaagg
     3061 catggaaaaa taccaaacca agaatagaga agttcagatc aagggcgggt acatgaaaat
     3121 agctaacgtt gggccaaaca ggatatctgc ggtgagcagt ttcggccccg gcccggggcc
     3181 aagaacagat ggtcaccgca gtttcggccc cggcccgagg ccaagaacag atggtcccca
     3241 gatatggccc aaccctcagc agtttcttaa gacccatcag atgtttccag gctcccccaa
     3301 ggacctgaaa tgaccctgcg ccttatttga attaaccaat cagcctgctt ctcgcttctg
     3361 ttcgcgcgct tctgcttccc gagctctata aaagagctca caacccctca ctcggcgcgc
     3421 cagtcctccg acagactgag tcgcccgggg gtctagtgcc accatgaccg agtacaagcc
     3481 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     3541 gttcgccgac taccccgcca cgcgccacac cgtggacccg gaccgccaca tcgagcgggt
     3601 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     3661 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     3721 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     3781 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     3841 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     3901 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     3961 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4021 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc atgcagatct ttgtgaagac
     4081 cctcactggc aaaaccatca cccttgaggt cgagcccagt gacaccattg agaatgtcaa
     4141 agccaaaatt caagacaagg agggtatccc acctgaccag cagcgtctga tatttgccgg
     4201 cagacagctg gaggatggcc gcactctctc agactacaac atccagaaag agtccaccct
     4261 gcacctggtg ttgcgcctgc gaggtggcat gggttaccca tacgatgttc cagattacgc
     4321 ttctagagga tccgcggccg ccttcttaac ccaacagaag gctcgagaag gtatattgct
     4381 gttgacagtg agcgcacaaa cgctctcatc gacaagtagt gaagccacag atgtacttgt
     4441 cgatgagagc gtttgtatgc ctactgcctc ggacttcaag gggctagaat tcggcagctg
     4501 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaact gcatccggac
     4561 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     4621 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     4681 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4741 tctagcaggg cccgtttcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5221 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5701 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6541 cgcacatttc cccgaaaagt gccacctgac
//
