LOCUS       V046000                 9284 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046000
VERSION     V046000
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    2058..2175
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        2227..2530
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2531..2734
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2826..2915
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             2931..4445
                     /label="PFKFB2(NM_006212)"
                     /note="PFKFB2(NM_006212)"
                     /gene="PFKFB2"
     CDS             4452..4523
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     promoter        4541..5040
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5061..5660
                     /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    5674..6262
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6337..6570
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        6775..6879
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6880..7740
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      7911..8499
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    8910..8931
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        8946..8976
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8984..9000
                     /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"
     primer_bind     9008..9024
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
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 tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccac gcgtgcggcc
     2821 gcatggagca gaaactcatc tctgaagaag atctggaaca aaagttgatt tcagaagaag
     2881 atctggaaca gaagctcatc tctgaggaag atctggcccg ggcgggatcc atgtctgggg
     2941 catcttcctc agaacagaac aacaacagct atgaaaccaa aaccccaaat cttcgaatgt
     3001 cagagaagaa atgttcatgg gcctcctaca tgaccaactc cccgactctg atcgttatga
     3061 ttggtttgcc agcccggggt aaaacctacg tgtccaagaa actaacacgc tacctcaact
     3121 ggattggagt ccccaccaaa gtgtttaatc ttggggtgta tcggcgtgaa gcagtcaagt
     3181 cctataagtc ctacgacttc tttcggcatg acaatgagga ggccatgaag atccgcaaac
     3241 agtgtgctct ggtggcgctg gaagatgtta aggcgtatct cactgaggag aatggtcaga
     3301 ttgcggtgtt tgatgccacc aatacaaccc gggagaggag ggacatgatt ttgaactttg
     3361 ctgaacagaa ttccttcaag gtattctttg tggaatccgt ctgtgatgat cctgatgtca
     3421 ttgctgccaa tattctggag gttaaggtat caagccctga ctatcctgaa aggaacagag
     3481 agaacgtgat ggaggacttc ctgaagagaa ttgaatgcta caaagttacc taccgacctc
     3541 ttgacccaga caactatgac aaggatcttt ctttcatcaa ggtgataaac gtgggccagc
     3601 gatttttagt caacagagtc caggactaca tccagagcaa gatagtctac tacctcatga
     3661 atatccacgt ccagcctcgc accatttacc tttgccggca tggagaaagc gagttcaatc
     3721 tcttggggaa gattgggggt gactctggcc tctcggtgcg gggaaagcag tttgcccaag
     3781 ctctaaggaa atttctggag gaacaggaaa taacagacct caaagtgtgg acaagccagt
     3841 tgaagaggac catacagact gctgaatctc tcggggtgcc ctatgagcag tggaagattc
     3901 tgaatgagat tgatgctggt gtgtgtgaag agatgaccta tgcagagatt gagaaacggt
     3961 acccagaaga gtttgcactt cgagatcaag agaagtatct gtatcgatat cctggtgggg
     4021 agtcatacca ggacctggtg cagcggctgg agcctgtcat catggagctg gaacgtcagg
     4081 gcaatgtcct cgtcatctcc caccaggctg tcatgcgctg cctcctggcc tacttcttgg
     4141 ataagggcgc agatgagcta ccatacttga gatgccctct ccataccatc ttcaaactta
     4201 ctcctgtggc ctatgggtgc aaagtggaaa caattaaact taacgtggag gctgtgaaca
     4261 cgcaccgtga caagccaact aacaacttcc ccaagaacca aacccctgta aggatgagaa
     4321 ggaacagctt tacgcctctg tccagttcga atacaataag gcgtccaaga aattacagtg
     4381 ttgggagccg gcccctcaag cccctcagcc ctctccgtgc ccaggacatg caagaagggg
     4441 ccgacctcga ggattacaag gatgacgacg ataaggacta taaggacgat gatgacaagg
     4501 actacaaaga tgatgacgat aaatagtcga gaattctacc gggtagggga ggcgcttttc
     4561 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     4621 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     4681 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     4741 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     4801 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     4861 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     4921 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     4981 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     5041 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     5101 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     5161 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     5221 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     5281 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     5341 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     5401 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5461 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5521 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5581 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5641 cccgcaagcc cggtgcctga atcgatagat cctaatcaac ctctggatta caaaatttgt
     5701 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5761 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5821 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5881 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5941 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     6001 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6061 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     6121 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6181 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6241 tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg
     6301 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6361 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6421 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6481 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6541 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6601 atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa
     6661 gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6721 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7081 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     7141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7561 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     8101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8581 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8641 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     8701 cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt
     8761 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     8821 gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac
     8881 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8941 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     9001 tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca
     9061 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
     9121 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
     9181 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
     9241 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt
//
