LOCUS       V031433                 9831 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031433
VERSION     V031433
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             2827..4557
                     /label="CSTF2(NM_001325)"
                     /note="CSTF2(NM_001325)"
                     /gene="CSTF2"
     CDS             4564..4587
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4619..5118
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5139..5738
                     /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    5752..6340
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6547..7180
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7309..7325)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7333..7349
                     /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(7357..7387)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7402..7423
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7711..8299)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8470..9330)
                     /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(9331..9435)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9483..9617
                     /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 gccaccatgg cgggtttgac tgtgagagac ccagcggtgg atcgttctct acgttctgtg
     2881 ttcgtgggga acattcctta tgaagctact gaagagcagt tgaaggacat cttttctgag
     2941 gttggacctg ttgttagttt cagattggta tacgatagag agacaggaaa gccaaagggt
     3001 tatggcttct gtgaatacca agaccaagag acagcactta gtgccatgcg gaacctgaat
     3061 gggcgcgaat tcagtgggag agcacttcga gtggacaatg ctgccagtga aaagaacaaa
     3121 gaagagctga agagccttgg cactggtgcc cctgtcattg agtcacctta tggagagacc
     3181 atcagtcctg aggatgcccc tgagtccatt agcaaagcag ttgccagcct tccaccagag
     3241 cagatgtttg agctgatgaa acaaatgaag ctctgtgtcc agaatagtcc ccaggaggca
     3301 cggaacatgt tacttcagaa ccctcaactg gcttatgctt tgctgcaagc acaggtagtg
     3361 atgagaattg tggatccgga aattgccctg aaaattctgc atcgccagac aaatatccca
     3421 acgctgattg caggcaaccc tcagccagtc catggtgctg ggcctggctc aggatccaat
     3481 gtgtcaatga accagcagaa tcctcaggcc cctcaggccc agtctttggg tggaatgcat
     3541 gtcaatggcg cacctcctct gatgcaagct tctatgcagg gtggagttcc agcaccaggg
     3601 caaatgccag ctgctgtcac aggacctggc cctggttcct tagctcctgg aggaggaatg
     3661 caggctcagg ttggaatgcc aggaagtgga ccagtgtcca tggaacgggg gcaagtgccg
     3721 atgcaagacc ccagagcagc tatgcagcgg ggatccttgc ctgcgaatgt cccaacccct
     3781 cgaggcttgt taggagatgc tccgaatgat ccacggggag gcactttact ttctgtaact
     3841 ggagaggtag agcctagagg ttacttggga ccacctcatc agggtccacc catgcaccat
     3901 gtccctggcc atgagagccg aggaccaccc ccacatgaac tgaggggagg gccattaccc
     3961 gagcccagac ctctaatggc agaaccaaga ggacccatgc tagatcagag gggtccaccc
     4021 ttggatggca gaggtggaag ggatccccga ggaatagatg cacgagggat ggaggcccga
     4081 gccatggagg caagagggtt agatgccaga ggattagagg cccgtgcaat ggaggcccgt
     4141 gcgatggaag ctcgtgcaat ggaggcccga gcgatggagg cccgtgcaat ggaagtccga
     4201 gggatggagg ccagaggcat ggataccaga ggcccagtgc ctggccccag aggacctata
     4261 cctagtggaa tgcagggtcc cagtccaatt aacatggggg cggttgtccc ccagggatcc
     4321 agacaggtcc cagtcatgca gggaacagga atgcaaggag caagtataca gggtggaagc
     4381 cagcctggcg gctttagtcc cgggcagaac caagtcactc cacaggatca tgagaaggct
     4441 gctttgatta tgcaggttct acaactgact gcagaccaga ttgccatgtt gcctcctgag
     4501 caaaggcaga gtatcctgat tttaaaggaa caaatacaga aatccactgg agcacctgaa
     4561 ttcgattaca aggatgacga cgataagtga ggatcccgcg actctagata attctaccgg
     4621 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     4681 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     4741 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     4801 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     4861 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     4921 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     4981 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     5041 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     5101 tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac aagcccacgg
     5161 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5221 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5281 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5341 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5401 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5461 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5521 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5581 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5641 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5701 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa caatcaacct
     5761 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5821 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5881 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5941 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6001 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6061 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6121 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     6181 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6241 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     6301 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc
     6361 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     6421 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     6481 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga
     6541 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     6601 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat
     6661 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag
     6721 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg
     6781 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg
     6841 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac
     6901 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc
     6961 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag
     7021 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     7081 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7141 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     7201 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat
     7261 tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     7321 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7381 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7441 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7501 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7561 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7621 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7681 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7741 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7801 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7861 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7921 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7981 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     8041 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     8101 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     8161 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8221 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8281 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8341 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8521 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8581 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8641 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8701 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8761 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8821 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8881 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8941 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9001 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9061 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9121 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9181 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9241 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9301 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9361 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9421 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9481 tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     9541 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     9601 cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca
     9661 ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt
     9721 cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag
     9781 tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag t
//
