LOCUS       V021118                 9743 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021118
VERSION     V021118
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"
     regulatory      2809..2818
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2818..2883
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2890..4434
                     /label="Trim35(NM_001025142)"
                     /note="Trim35(NM_001025142)"
                     /gene="Trim35"
     misc_feature    4446..5032
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5052..5651
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5665..6253
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6460..7093
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7221..7237)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7245..7261
                     /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(7269..7299)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7314..7335
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7623..8211)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8382..9242)
                     /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(9243..9347)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9395..9529
                     /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 caccatggac
     2821 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     2881 aagctcgaga tgaaggcggc tgcatcggtg gtgacggctg ctgctcctgc gatggagcca
     2941 ggcccctctg tgtcccccgg gccttcgcgc tccttcaaag aggagctgct gtgtgccgtg
     3001 tgctacgacc cgttccgcga cgcagtgact ctgcgctgtg gccacaactt ctgccgccgg
     3061 tgcgtgagcg gctgctggga ggtgcagacg acgccctcgt gcccggtgtg caaggaacga
     3121 gcggtgcccg gggagctgcg caccaaccac acgctcaaca acctggtgga gaccttgctg
     3181 cgcgaggagg ccgagggcgc gcgctggacc ggtcgccggt ccccgcgccc ctgccgcgcg
     3241 caccgtgccc cgctcacgct cttctgcgtg gaggacaagg agctgttgtg ctgtgcctgt
     3301 caagccgacg cccggcacca ggagcatcgt gtgcaaccca tcaaggacac tgcgcaggac
     3361 ttccgggcca agtgtaagaa catggagcac gtacttcggg agaaagccaa gtccttctgg
     3421 gctctgaggc gcacctatga ggccattgcc aagcacaatg aggtgcagac aacttggcta
     3481 gaaggccgaa tccgggatga gtttgacaag ctccgagatt tcctgagggt ggaggaacaa
     3541 gccaccgtgg atgccatgaa ggaagagagc aggaagaagc acctgctggc tgaggagaag
     3601 atgaagcagc tggcggagca gaccgaggca ctggctcgag agattgagcg tctgcagatg
     3661 gagatgaagg aagacgacat gaccttcctc atgaaacaca agagccgaaa acgccggctc
     3721 ttctgcaccg tggagccagc acctctccag cctggcttgc ttatggacgc atgcaagtat
     3781 ctggagtcct tgcagtaccg agtctggaag aagatgcttg ggtctgttga atcggtgccc
     3841 ttcagcttgg atcccaacac agctgctggc tggctcaaag tggctgatga cctcacgagc
     3901 gtcatcaacc atggctaccg tgtgcaggtg gagaatccag agcgcttctc ctcggcaccc
     3961 tgcctgctgg gctctcaagt tttctccaag ggctcccact cttgggaggt ggatgtgggt
     4021 ggcctgccga gctggcgagt gggcgtggtg cgggtgcagg cacatgcaca ggcacaggct
     4081 caggctgatg tcggtggtga gggccactca cacagctgtt accatgatac acgctcaggc
     4141 ttctggtacc tgtgccgcac tcagggtgtg gacggagacc actgcatgac ttccgacaca
     4201 gccacagccc ctctggtcca ggccatgcca cgccgtctgc gtgtggagct ggagtgtgag
     4261 gagggtgagc tgtccttcta tgactctgag cgccactgcc atctgtatac cttccatgcc
     4321 cactttgggg aggtgaggcc ctacttctac ctaggggcct ctcggggtga tggtccccca
     4381 gaacctctgc gcatctgcca cctgcgtgtc tccatcaaag aagagctgga catctgagga
     4441 tcccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     4501 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     4561 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     4621 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     4681 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     4741 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     4801 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     4861 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     4921 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     4981 cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga
     5041 gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5101 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5161 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5221 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5281 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5341 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5401 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5461 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5521 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5581 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5641 ccggtgccta gacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg
     5701 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5761 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5821 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5881 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5941 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6001 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     6061 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6121 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6181 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6241 gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca
     6301 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     6361 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     6421 ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc
     6481 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     6541 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
     6601 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
     6661 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
     6721 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
     6781 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
     6841 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
     6901 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6961 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7021 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7081 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7141 aaatgaatat cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc
     7201 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     8101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9361 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg
     9421 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     9481 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
     9541 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
     9601 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     9661 ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg tatttgttaa
     9721 atatgtacta caaacttagt agt
//
