LOCUS       V021415                 8658 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021415
VERSION     V021415
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..3309
                     /label="ISG15(NM_005101)"
                     /note="ISG15(NM_005101)"
                     /gene="ISG15"
     misc_feature    3329..3901
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             3903..4613
                     /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    4627..5215
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5422..6055
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6183..6199)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6207..6223
                     /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(6231..6261)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6276..6297
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6585..7173)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7344..8204)
                     /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(8205..8309)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8357..8491
                     /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 caccatgggc
     2821 tgggacctga cggtgaagat gctggcgggc aacgaattcc aggtgtccct gagcagctcc
     2881 atgtcggtgt cagagctgaa ggcgcagatc acccagaaga tcggcgtgca cgccttccag
     2941 cagcgtctgg ctgtccaccc gagcggtgtg gcgctgcagg acagggtccc ccttgccagc
     3001 cagggcctgg gccccggcag cacggtcctg ctggtggtgg acaaatgcga cgaacctctg
     3061 agcatcctgg tgaggaataa caagggccgc agcagcacct acgaggtacg gctgacgcag
     3121 accgtggccc acctgaagca gcaagtgagc gggctggagg gtgtgcagga cgacctgttc
     3181 tggctgacct tcgaggggaa gcccctggag gaccagctcc cgctggggga gtacggcctc
     3241 aagcccctga gcaccgtgtt catgaatctg cgcctgcggg gaggcggcac agagcctggc
     3301 gggcggagct aagcggccgc ggatcccgcc cctctccctc ccccccccct aacgttactg
     3361 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     3421 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     3481 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     3541 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     3601 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     3661 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     3721 aatggctcac ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     3781 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     3841 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     3901 atatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     3961 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     4021 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     4081 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     4141 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     4201 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     4261 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     4321 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     4381 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     4441 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     4501 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     4561 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tgaacgcgtc
     4621 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4681 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4741 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4801 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4861 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4921 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4981 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     5041 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5101 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5161 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     5221 attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     5281 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     5341 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     5401 ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc
     5461 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
     5521 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     5581 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
     5641 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
     5701 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
     5761 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
     5821 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
     5881 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5941 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6001 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6061 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6121 gagaggcctt gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca
     6181 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6241 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6301 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6361 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6421 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6481 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6541 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6781 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7021 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7621 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8341 gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8401 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8461 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
     8521 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
     8581 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
     8641 tactacaaac ttagtagt
//
