LOCUS       V021579                10523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021579
VERSION     V021579
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             2826..5219
                     /label="Itgb1(NM_010578)"
                     /note="Itgb1(NM_010578)"
                     /gene="Itgb1"
     CDS             5226..5291
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5311..5810
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5831..6430
                     /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    6444..7032
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7239..7872
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(8001..8017)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8025..8041
                     /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(8049..8079)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8094..8115
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8403..8991)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9162..10022)
                     /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(10023..10127)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10175..10309
                     /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 ccaccatgaa tttgcaactg gtttcctgga ttggattgat cagtttgatt tgttctgtat
     2881 ttggccaaac agataaaaat agatgtttaa aagcaaatgc caaatcttgc ggagaatgta
     2941 tacaagcagg gccaaattgt gggtggtgta caaatacgac atttttgcaa gaaggaatgc
     3001 ctacttctgc acgatgtgat gatttagaag ctttgaaaaa gaagggttgc cagccaagtg
     3061 acatagagaa tcccagaggc tctcaaacta taaagaaaaa taaaaatgtc accaatcgca
     3121 gcaaagggat ggcagagaag ctccggccag aagacattac tcagatccaa ccacaacagc
     3181 tgcttctaaa attgagatca ggagaaccac agaagtttac attaaaattc aagagggctg
     3241 aagattaccc tattgatctc tactacctta tggatctctc ctactctatg aaagatgatc
     3301 tggagaatgt gaaaagtctt ggaacggatt tgatgaatga aatgaggagg attacttcag
     3361 acttccgcat tggctttggc tcatttgtgg agaaaactgt gatgccgtat attagcacaa
     3421 ccccagcaaa gctaagaaat ccttgtacaa gtgaacaaaa ctgcaccagc ccatttagct
     3481 acaaaaatgt gcttagtctt actgacagag gagagttttt caatgaactt gttggtcagc
     3541 aacgcatatc tggaaacttg gattctccag aaggtggctt tgatgcaatc atgcaggttg
     3601 cggtttgtgg atcgctgatt ggctggagga atgtaacacg actgctggtg ttttccacgg
     3661 atgctgggtt tcactttgct ggagatggga aacttggtgg tattgtttta cccaatgatg
     3721 gacaatgtca cctggaaaat aatgtatata caatgagcca ttactatgat tatccttcaa
     3781 ttgctcacct tgttcagaaa ctaagtgaaa ataatattca gacgattttt gcagttactg
     3841 aagagttcca acctgtttac aaggaattga agaatttgat tcctaagtca gcagtgggca
     3901 cactgtctgg aaactctagt aatgtgatcc agctaatcat cgatgcctac aactctcttt
     3961 cttcagaagt cattctggaa aatagcaaat tgccagacgg agtaacaata aattacaaat
     4021 cctattgcaa gaatggggtg aatgggacag gagaaaatgg acgaaagtgt tccaacattt
     4081 ctattggaga tgaggttcaa tttgaaatta gcataactgc taataaatgt ccaaataagg
     4141 agtctgaaac cattaaaatt aaacctctgg gcttcactga agaagtagag gtcgttcttc
     4201 agttcatctg taagtgcaat tgtcaaagcc atggcatccc agccagtccc aagtgccatg
     4261 agggaaatgg gacatttgag tgtggagcct gcaggtgcaa tgaggggcgt gttgggaggc
     4321 actgtgaatg tagcacagat gaagtgaaca gtgaagacat ggacgcttac tgcaggaaag
     4381 agaacagttc ggaaatctgc agtaacaatg gagaatgtgt ctgtggacag tgtgtgtgta
     4441 ggaagagaga taatacaaat gaaatttact ctggaaaatt ctgcgagtgt gataacttca
     4501 actgtgatag gtctaatggc ttaatttgtg gaggcaatgg cgtgtgcagg tgtcgtgttt
     4561 gtgaatgcta tcccaattac actggcagtg catgtgactg ttctttggac actggtccat
     4621 gtctagcgtc aaatggtcag atctgcaatg gccggggtat ttgtgaatgt ggtgcttgta
     4681 agtgcacaga tcccaagttt caagggccaa cttgtgagac atgtcagacc tgccttggcg
     4741 tctgtgcaga gcataaagaa tgtgttcagt gcagagcctt caataaagga gaaaagaaag
     4801 acacgtgtgc acaggagtgc tcccacttca atctcaccaa agtagaaagc agggagaagt
     4861 tgccccagcc ggtgcaggtc gatcctgtga cccattgcaa ggagaaggac attgatgact
     4921 gctggttcta tttcacctat tcagtgaatg gcaacaatga agctatcgtg catgttgtgg
     4981 agactccaga ctgtcctact ggtcccgaca tcatcccaat tgtagcaggc gtggttgctg
     5041 gaattgttct tattggcctt gccttgctgc tgatttggaa acttttaatg ataattcatg
     5101 acagaaggga atttgctaaa tttgaaaagg agaaaatgaa tgccaagtgg gacacgggtg
     5161 aaaatcctat ttacaagagt gccgtgacaa ctgtggtcaa tccgaagtat gagggaaaag
     5221 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     5281 acgatgacaa gtagtctaga taattctacc gggtagggga ggcgcttttc ccaaggcagt
     5341 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     5401 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     5461 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     5521 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     5581 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     5641 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     5701 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     5761 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     5821 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5881 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5941 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     6001 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     6061 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     6121 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     6181 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     6241 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     6301 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     6361 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     6421 cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     6481 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6541 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6601 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6661 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6721 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6781 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6841 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6901 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6961 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     7021 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     7081 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     7141 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     7201 tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca
     7261 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta
     7321 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
     7381 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
     7441 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag
     7501 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa
     7561 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg
     7621 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
     7681 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     7741 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7801 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7861 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7921 aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc gacctctagc
     7981 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     8041 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     8101 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     8161 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     8221 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     8281 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     8341 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     8401 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     8461 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     8521 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8581 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8641 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8701 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8761 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8821 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     8881 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8941 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     9001 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     9061 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     9121 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     9181 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     9241 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     9301 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     9361 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     9421 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     9481 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9541 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9601 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9661 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9721 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9781 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9841 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9901 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9961 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    10021 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    10081 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    10141 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg
    10201 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    10261 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
    10321 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
    10381 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
    10441 ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg tatttgttaa
    10501 atatgtacta caaacttagt agt
//
