LOCUS       V035617                 9983 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035617
VERSION     V035617
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             2829..2894
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2901..4739
                     /label="IGSF8(NM_052868)"
                     /note="IGSF8(NM_052868)"
                     /gene="IGSF8"
     promoter        4771..5270
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5291..5890
                     /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    5904..6492
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6699..7332
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7461..7477)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7485..7501
                     /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(7509..7539)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7554..7575
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7863..8451)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8622..9482)
                     /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(9483..9587)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9635..9769
                     /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 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2881 atgacgatga caaggaattc atgggcgccc tcaggcccac gctgctgccg ccttcgctgc
     2941 cgctgctgct gctgctaatg ctaggaatgg gatgctgggc ccgggaggtg ctggtccccg
     3001 aggggccctt gtaccgcgtg gctggcacag ctgtctccat ctcctgcaat gtgaccggct
     3061 atgagggccc tgcccagcag aacttcgagt ggttcctgta taggcccgag gccccagata
     3121 ctgcactggg cattgtcagt accaaggata cccagttctc ctatgctgtc ttcaagtccc
     3181 gagtggtggc gggtgaggtg caggtgcagc gcctacaagg tgatgccgtg gtgctcaaga
     3241 ttgcccgcct gcaggcccag gatgccggca tttatgagtg ccacaccccc tccactgata
     3301 cccgctacct gggcagctac agcggcaagg tggagctgag agttcttcca gatgtcctcc
     3361 aggtgtctgc tgccccccca gggccccgag gccgccaggc cccaacctca cccccacgca
     3421 tgacggtgca tgaggggcag gagctggcac tgggctgcct ggcgaggaca agcacacaga
     3481 agcacacaca cctggcagtg tcctttgggc gatctgtgcc cgaggcacca gttgggcggt
     3541 caactctgca ggaagtggtg ggaatccggt cagacttggc cgtggaggct ggagctccct
     3601 atgctgagcg attggctgca ggggagcttc gtctgggcaa ggaagggacc gatcggtacc
     3661 gcatggtagt agggggtgcc caggcagggg acgcaggcac ctaccactgc actgccgctg
     3721 agtggattca ggatcctgat ggcagctggg cccagattgc agagaaaagg gccgtcctgg
     3781 cccacgtgga tgtgcagacg ctgtccagcc agctggcagt gacagtgggg cctggtgaac
     3841 gtcggatcgg cccaggggag cccttggaac tgctgtgcaa tgtgtcaggg gcacttcccc
     3901 cagcaggccg tcatgctgca tactctgtag gttgggagat ggcacctgcg ggggcacctg
     3961 ggcccggccg cctggtagcc cagctggaca cagagggtgt gggcagcctg ggccctggct
     4021 atgagggccg acacattgcc atggagaagg tggcatccag aacataccgg ctacggctag
     4081 aggctgccag gcctggtgat gcgggcacct accgctgcct cgccaaagcc tatgttcgag
     4141 ggtctgggac ccggcttcgt gaagcagcca gtgcccgttc ccggcctctc cctgtacatg
     4201 tgcgggagga aggtgtggtg ctggaggctg tggcatggct agcaggaggc acagtgtacc
     4261 gcggggagac tgcctccctg ctgtgcaaca tctctgtgcg gggtggcccc ccaggactgc
     4321 ggctggccgc cagctggtgg gtggagcgac cagaggacgg agagctcagc tctgtccctg
     4381 cccagctggt gggtggcgta ggccaggatg gtgtggcaga gctgggagtc cggcctggag
     4441 gaggccctgt cagcgtagag ctggtggggc cccgaagcca tcggctgaga ctacacagct
     4501 tggggcccga ggatgaaggc gtgtaccact gtgcccccag cgcctgggtg cagcatgccg
     4561 actacagctg gtaccaggcg ggcagtgccc gctcagggcc tgttacagtc tacccctaca
     4621 tgcatgccct ggacacccta tttgtgcctc tgctggtggg tacaggggtg gccctagtca
     4681 ctggtgccac tgtccttggt accatcactt gctgcttcat gaagaggctt cgaaaacggt
     4741 gaggatcccg cgactctaga taattctacc gggtagggga ggcgcttttc ccaaggcagt
     4801 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     4861 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     4921 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     4981 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     5041 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     5101 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     5161 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     5221 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     5281 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5341 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5401 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5461 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5521 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5581 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     5641 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     5701 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     5761 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     5821 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     5881 cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     5941 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6001 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6061 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6121 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6181 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6241 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6301 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6361 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6421 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6481 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     6541 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     6601 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     6661 tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca
     6721 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta
     6781 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
     6841 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
     6901 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag
     6961 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa
     7021 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg
     7081 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
     7141 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     7201 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7261 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7321 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7381 aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc gacctctagc
     7441 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7501 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7561 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7621 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7681 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8041 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8281 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     8341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8641 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8701 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8761 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8821 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8881 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8941 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9001 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9061 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9121 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9181 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9241 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9301 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9361 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9421 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9481 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9541 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9601 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg
     9661 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     9721 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
     9781 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
     9841 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     9901 ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg tatttgttaa
     9961 atatgtacta caaacttagt agt
//
