LOCUS       V021652                 9641 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021652
VERSION     V021652
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      2820..2829
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2835..2924
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             2958..4397
                     /label="GRSF1(NM_002092)"
                     /note="GRSF1(NM_002092)"
                     /gene="GRSF1"
     promoter        4429..4928
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4949..5548
                     /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    5562..6150
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6357..6990
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7119..7135)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7143..7159
                     /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(7167..7197)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7212..7233
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7521..8109)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8280..9140)
                     /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(9141..9245)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9293..9427
                     /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 tcgctagcgc taccggactc agatctcgag
     2821 ccaccatggc ggcggaacaa aaactaatat cagaggaaga tttggaacag aaactaatca
     2881 gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttatcagcg gaaaagcttg
     2941 gtaccgagct cggatccatg gccggcacgc gctgggtact cggggcgctg ctccggggct
     3001 gcggctgtaa ctgcagcagc tgccggcgca ccggcgccgc ctgcctgccc ttctactccg
     3061 ccgctggctc tatcccgtcg ggcgtctcgg gccgccgccg cctgctgctg ctgctcgggg
     3121 ccgccgcggc cgctgcctcc cagacgcgtg gcctccagac cgggcctgtg cctcccggga
     3181 ggctggcggg gcctcccgct gtggccacct ctgccgcggc cgcggccgcc gcgtcctact
     3241 ctgccctccg tgcctctctg ctgccgcagt cgctggcggc ggcggccgcc gtcccgacgc
     3301 gcagctacag ccaggagtcc aaaactactt acctggaaga ccttccacca ccccctgagt
     3361 atgaattggc cccgtccaag ttagaagagg aagtggatga tgtctttctc attcgagctc
     3421 aaggactgcc ctggtcatgc actatggaag atgtgcttaa ctttttttca gactgcagaa
     3481 tccgcaacgg tgagaatgga atacattttc tcctaaacag agatgggaaa cgaaggggtg
     3541 atgccttaat tgaaatggag tcagagcagg atgtgcagaa agccttagag aagcaccgca
     3601 tgtacatggg ccagcggtat gtggaagtat atgagataaa caatgaagat gtggatgcct
     3661 taatgaagag cttgcaggtc aaatcttcgc ctgtggtaaa tgatggtgtg gttcgtttga
     3721 gaggacttcc ttatagttgc aatgagaaag acattgtaga cttctttgca ggactgaata
     3781 tagttgacat tacttttgtg atggactata gagggaggcg aaaaacaggg gaagcctatg
     3841 tgcaatttga agaaccagaa atggccaacc aagccctgtt gaaacacagg gaagaaattg
     3901 gtaatcgata catcgagata tttccaagca gaaggaatga agttcgaaca catgtcggtt
     3961 cttataaggg aaagaaaatc gcatcttttc ctactgctaa gtatataact gagccagaaa
     4021 tggtctttga agaacatgaa gtaaatgagg atattcaacc catgacagct tttgaaagtg
     4081 agaaggaaat agaattgcct aaggaggtgc cagaaaagct tccagaggct gctgattttg
     4141 gaactacgtc ttctctgcat tttgtccaca tgagaggatt acctttccaa gccaatgccc
     4201 aagacattat aaactttttt gctccactca agcctgttag aatcaccatg gaatacagct
     4261 ccagtgggaa ggccactgga gaagctgatg tgcactttga gacccatgag gatgctgttg
     4321 cagcgatgct caaggatcgg tcccacgttc atcataggta tattgaactg ttcctgaatt
     4381 catgtccaaa aggaaaataa ggatcccgcg actctagata attctaccgg gtaggggagg
     4441 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     4501 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     4561 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     4621 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
     4681 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     4741 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     4801 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     4861 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     4921 gcctttcgac ctgcagccca agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     4981 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5041 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5101 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5161 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5221 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5281 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5341 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5401 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5461 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5521 gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa caatcaacct ctggattaca
     5581 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5641 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5701 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5761 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5821 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5881 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5941 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     6001 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6061 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6121 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga
     6181 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     6241 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
     6301 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga
     6361 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
     6421 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
     6481 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
     6541 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
     6601 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
     6661 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
     6721 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
     6781 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     6841 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6901 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6961 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     7021 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt
     7081 ttaccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     7141 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7201 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7321 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7381 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7441 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7501 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7561 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7621 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7681 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7741 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7801 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7861 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7921 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7981 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8041 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8101 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8281 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8341 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8401 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8461 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8521 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8581 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8641 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8701 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8761 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8821 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8881 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8941 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9001 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9061 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9121 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9181 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9241 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt
     9301 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     9361 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     9421 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
     9481 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
     9541 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagtttttaa
     9601 agaaattgta tttgttaaat atgtactaca aacttagtag t
//
