LOCUS       V021395                 9372 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021395
VERSION     V021395
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..4023
                     /label="Trim59(NM_025863)"
                     /note="Trim59(NM_025863)"
                     /gene="Trim59"
     misc_feature    4043..4615
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4617..5327
                     /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    5341..5929
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6136..6769
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6897..6913)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6921..6937
                     /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(6945..6975)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6990..7011
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7299..7887)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8058..8918)
                     /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(8919..9023)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9071..9205
                     /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 caccatgcac
     2821 aattttgagg aggagttaac gtgtcccatc tgttacagca tctttgaaga tccccgtgta
     2881 ctaccatgct ctcatacatt ttgtagaaac tgtttggaaa atgttcttca ggcatctggt
     2941 aacttttaca tatggaggcc tctgcgaatt ccactcaagt gtcctaactg cagaagtatt
     3001 attgaaattg cttctactgg catagaatcc ttacctgtaa actttgcatt acgagcaatt
     3061 attgaaaagt accagcaaga agaccaccca gatgttgtca cctgccctga acactacagg
     3121 caaccattaa atgtttattg tctactagat aagaaattag tttgtggcca ttgtcttact
     3181 ataggccaac atcatggcca tcctatagat gaccttcaaa gtgcctatct gaaagaaaag
     3241 gatacacctc agaagttgtt gaaacagtta actgacacac attggacaga tatcactcgc
     3301 cttattgaaa agcttgaaga acagaaatgt cattctgaga aaatagttca aggtgataag
     3361 gaagttgttc tccagtattt taaggagcta attgatacat tagaacagaa aaaaaagtat
     3421 tttttggcag ctctttgtga tgttggcaaa atgattaatc aagaatacac tccacagatt
     3481 caaggaatga aggagataag ggaacaacaa cttgaattaa tgacaataac gacatcttta
     3541 caagatgagt ctccacttaa atttctggaa aaaattgatg aagtccgcca acgtgtgcag
     3601 atgttgaaac agagaccact tcctgaggtc cagcctgttg aaatttatcc tcgagtaagc
     3661 aatgtattaa aagaagagtg gagcagaata gaaattggac gtattaagaa agctgttatt
     3721 cctgaaatga gagtttcttc aaaaagaacg ccatgttcct ggtctgataa tgatgaaaaa
     3781 gaaatggaac tttttaaaat tttaaacatt gctatagttt cactcatttc agtgatacta
     3841 atgttgatac tccttttcaa ccatcacata ataacattct taaatgaaat cacttcaata
     3901 tgtttctctg aagtctttct ctctgtttac cagagtttat ctaagaactt atatgactta
     3961 aacaatacag tgtgttacac tttatatttg ttaaaggaat ttatgtggaa aatagtttct
     4021 cgttgagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     4081 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     4141 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     4201 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     4261 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     4321 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     4381 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     4441 tcacctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     4501 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg
     4561 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     4621 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4681 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4741 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4801 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4861 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4921 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     4981 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     5041 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     5101 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     5161 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     5221 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     5281 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaacg cgtctggaac
     5341 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5401 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5461 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5521 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5581 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5641 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5701 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     5761 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5821 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5881 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat
     5941 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     6001 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     6061 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     6121 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     6181 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
     6241 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
     6301 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
     6361 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
     6421 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
     6481 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
     6541 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
     6601 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6661 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6721 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6781 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6841 ccttgacatt gctagcgttt accgtcgacc tctagctaga gcttggcgta atcatggtca
     6901 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6961 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7021 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7081 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7141 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7201 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7261 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7321 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7381 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7441 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7501 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7561 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7621 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7681 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7741 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7801 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7981 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8041 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8101 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8161 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8221 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8281 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8341 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8401 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8461 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8521 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8581 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8641 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8701 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8761 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8821 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8881 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8941 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9001 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9061 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9181 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     9241 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     9301 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     9361 aaacttagta gt
//
