LOCUS       V021378                 9279 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021378
VERSION     V021378
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2815..3930
                     /label="USP18(NM_017414)"
                     /note="USP18(NM_017414)"
                     /gene="USP18"
     misc_feature    3950..4522
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4524..5234
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    5248..5836
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6043..6676
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6804..6820)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6828..6844
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6852..6882)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6897..6918
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7206..7794)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7965..8825)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8826..8930)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8978..9112
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatgagc
     2821 aaggcgtttg ggctcctgag gcaaatctgt cagtccatcc tggctgagtc ctcgcagtcc
     2881 ccggcagatc ttgaagaaaa gaaggaagaa gacagcaaca tgaagagaga gcagcccaga
     2941 gagcgtccca gggcctggga ctaccctcat ggcctggttg gtttacacaa cattggacag
     3001 acctgctgcc ttaactcctt gattcaggtg ttcgtaatga atgtggactt caccaggata
     3061 ttgaagagga tcacggtgcc caggggagct gacgagcaga ggagaagcgt ccctttccag
     3121 atgcttctgc tgctggagaa gatgcaggac agccggcaga aagcagtgcg gcccctggag
     3181 ctggcctact gcctgcagaa gtgcaacgtg cccttgtttg tccaacatga tgctgcccaa
     3241 ctgtacctca aactctggaa cctgattaag gaccagatca ctgatgtgca cttggtggag
     3301 agactgcagg ccctgtatac gatccgggtg aaggactcct tgatttgcgt tgactgtgcc
     3361 atggagagta gcagaaacag cagcatgctc accctcccac tttctctttt tgatgtggac
     3421 tcaaagcccc tgaagacact ggaggacgcc ctgcactgct tcttccagcc cagggagtta
     3481 tcaagcaaaa gcaagtgctt ctgtgagaac tgtgggaaga agacccgtgg gaaacaggtc
     3541 ttgaagctga cccatttgcc ccagaccctg acaatccacc tcatgcgatt ctccatcagg
     3601 aattcacaga cgagaaagat ctgccactcc ctgtacttcc cccagagctt ggatttcagc
     3661 cagatccttc caatgaagcg agagtcttgt gatgctgagg agcagtctgg agggcagtat
     3721 gagctttttg ctgtgattgc gcacgtggga atggcagact ccggtcatta ctgtgtctac
     3781 atccggaatg ctgtggatgg aaaatggttc tgcttcaatg actccaatat ttgcttggtg
     3841 tcctgggaag acatccagtg tacctacgga aatcctaact accactggca ggaaactgca
     3901 tatcttctgg tttacatgaa gatggagtgc taagcggccg cggatcccgc ccctctccct
     3961 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     4021 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     4081 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     4141 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     4201 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     4261 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     4321 ggatagttgt ggaaagagtc aaatggctca cctcaagcgt attcaacaag gggctgaagg
     4381 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     4441 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     4501 cctttgaaaa acacgatgat aatatggtga gcaagggcga ggaggataac atggccatca
     4561 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     4621 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     4681 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     4741 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     4801 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     4861 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     4921 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     4981 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     5041 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     5101 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     5161 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     5221 agctgtacaa gtgaacgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     5281 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5341 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5401 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5461 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5521 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5581 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5641 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     5701 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     5761 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5821 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
     5881 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     5941 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     6001 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     6061 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     6121 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     6181 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     6241 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     6301 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     6361 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     6421 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     6481 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6541 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6601 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6661 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6721 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgtttacc gtcgacctct
     6781 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6841 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6901 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     7021 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7081 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7141 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7201 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7261 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7321 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7381 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7441 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7501 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7561 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7621 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7681 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7741 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7801 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7861 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7921 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7981 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8041 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8101 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8161 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8221 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8281 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8341 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8401 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8461 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8521 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8581 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8641 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8701 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8761 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8821 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8881 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8941 cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa
     9001 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     9061 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     9121 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     9181 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     9241 aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
